DEVELOPMENT OF FIBER AND MINERAL ENRICHED COOKIES BY UTILIZATION OF BANANA AND BANANA PEEL FLOUR
Md. Jahangir Alam*, Shamoli Akter, Shohana Afroze, Md. Tarikul Islam, Emdadul Haque Sayeem
Address(es): Department of Food Technology and Nutritional Science, Faculty of Life Science, Mawlana Bhashani Science and Technology University, Santosh, Tangail- 1902, Bangladesh.
*Corresponding author: [email protected], [email protected] doi: 10.15414/jmbfs.2020.10.3.329-334
ARTICLE INFO ABSTRACT
Received 7. 3. 2019 An experimental study was designed to formulate ready to eat cookies by incorporating banana and banana peel flour which is normally Revised 17. 5. 2020 unused in Bangladesh but contains excellent amount of nutrients especially dietary fiber, essential vitamins and minerals. Cookies were Accepted 30. 6. 2020 prepared by replacing 5 % (sample-1), 10 % (sample-2) and 15% (sample-3) of wheat flour with banana and banana peel flour. The Published 1. 12. 2020 proximate analysis and sensory parameters of those cookies were compared with control cookies where no banana and banana peel flour were added and designated as normal cookies (0% substitution). Functional properties were also evaluated and a significant difference found (P<0.05) in WHC, OHC, swelling capacity, emulsion activity, emulsion stability and flour dispersibility in banana peel flour Regular article when compared to wheat flour. On proximate analysis of cookies, significant variation (P<0.05) was also observed in protein, ash, fiber and carbohydrate content of banana and banana peel flour cookies in a comparison to normal cookies. The increasing the substitution of banana and banana peel flour in cookies increased the ash and crude fiber content remarkably. About 15% substitution of banana and banana peel flour in cookies increased 93.25% crude ash (mineral) and 197.56% crude fiber than normal cookies. Energy values of the cookies were also evaluated and ranged between 480 Kcal and 513 Kcal per 100 g, with sample-3 cookies having the lowest value. In conclusion, the addition of both banana and banana peel flour in cookies by replacing 10% wheat flour were more acceptable with all quality characteristics.
Keywords: Banana, cookies, dietary fiber, functional properties, sensory evaluation
INTRODUCTION a wing to produce functional food to replace traditionally commercialized products. With these grounds, peels of different fruits, vegetables and herbs Cookies are widely consumed as ready to eat and convenient foods throughout paying attention as they are rich source of natural dietary fiber and antioxidant. In the world which gives more nutrient than any other single food sources (Raihan this perspective, banana peel has attracted attention due to its rich source of and Saini, 2017). Now-a-days, cookies represent the leading category of snack dietary fiber (14.5%). Besides, banana peels are normally unused in south Asia foods in the major parts of the world (Laura and Eric, 2014). Traditionally, and also good sources of total starch (73.4%), resistant starch (17.5%), protein, cookies making processes are quite simple with some basic ingredients that fat and micronutrients (Juarez-Garcia et al., 2006). So that consumption of consist of flour, sugar and eggs. Generally, soft wheat is used to make cookies banana peels flour may play a significant role in human health (Rodriguez- (Laura, Eric and Emelem, 2013). Considering geographical scarceness, high Ambriz et al., 2008). New strategy in production of banana peel flour from cost and unavailibility of wheat flour, an alternative source of wheat flour are in unripe banana and incorporates it to different novel foods such as slowly high demand. Substation of wheat flour with other nutrient dense materials may digestible cookies (Aparicio-Saguilan et al., 2007) high-fiber and edible films reduce expense as well as import rate of wheat flour. This appeared the (Rungsinee and Natcharee, 2007). Several researchers improve the nutritional necessities of composite flour (Oyeku et al., 2008) which is defined as a mixture quality of cookies by incorporating composite flour from different types of food of whole wheat flour with non-wheat flours for the production of baking products sources (Chinma and Gernah, 2007; Singh, Riar and Saxena, 2008; Noor such as biscuits, cookies, and pastas (Lakshmi, Swarnali and Usha, 2015). Non- Aziah and Komathi, 2009). In this regard, this study was directed to assess the wheat flour (substitute) should be available, accessible, economical and effects of substituting different amount of nutritionally enriched banana and nutritionally enriched and able to fulfill the function of wheat flour (Akobundu, banana peel flour with wheat flour into cookies and evaluate its effect on texture, Ubbaonu and Ndupuh, 1998). It imparts an admirable means of increasing the sensory attributes and nutritional content. nutrient content like protein, essential vitamins, important minerals, and bioactive compounds of foods by incorporation of readily available non wheat flour MATERIALS & METHODS (Okafor, Ozumba and Solomon, 2002). This type of enrichment may play a significant role in developing countries like Bangladesh where malnutrition is Place of experiment prevalent. In this context banana and banana peel flour may be a great substitute of wheat flour because banana (Musa sapientum L) is one of the foremost fruit The sample analyses were conducted in the laboratory of Food Technology and crops in Bangladesh in respect of production and area (BBS, 2006). It is also an Nutritional Science (FTNS) Department, Mawlana Bhashani Science and economically considerable fruits in Bangladesh which is grown both in the Technology University (MBSTU), Santosh, Tangail-1902, Bangladesh. homestead and commercial farms. Banana is one of the cheapest, delicious and most nourishing of all fruits. It is very much popular for its characteristics aroma, Collection of raw materials texture and easy to peel and eat. Banana contain most of the essential vitamins including vitamin-A, C and B6, important minerals, good source of fat free The banana (Musa sapientam L), which local name is “Sagor kola” was dietary fiber and a rich source of calories (Hossain, Abdulla and Majumder, purchased from local market of Santosh, Tangail, Bangladesh. The other 2016). Good sources of dietary fiber is very important for human health, in this ingredients which were used in cookies production i.e. refined wheat flour, concept interests are increasing to find out new sources of dietary fibers that add hydrogenated vegetable oil, sugar powder, vegetable oil (soybean), egg, whole
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milk, iodized salt, baking powder and required chemicals were used from the Analysis of functional properties of banana peel and wheat flour laboratory of FTNS department, MBSTU and some of them procured from the local market. Functional qualities such as water and oil holding capacity, swelling capacity, emulsion activity and stability and also flour dispersibility of banana peel flour Preparation of banana (unripe) peel flour and wheat flour were analyzed.
At first the bananas were collected and washed with water to remove all harmful Water holding capacity (WHC) ingredients. After peeling, the peels were sliced and dipped for ten minutes in 0.5% (w/v) citric acid solution to reduce enzymatic reaction. After drained out, About 1g of flour sample and 25ml of water were taken in a 30 ml capacity test the sliced were then dried in oven at 500C for 16-20 hrs. The dried banana peels tube and allowed for 15 minutes at room temperature. The tube was then were then amalgamated by using a commercial blender and passes through 60 centrifuge for 20 min at 3000 rpm and drained off the supernatant. The weight of mesh screen to obtain fine banana peel flour. The dried banana peels powder the residue after drained off the excess water was the water holding capacity of were then stored in airtight packet at ambient temperature. flour and determined as g of water/g dry sample (Rodriguez-Ambriz et al., 2008). Formulation of cookies Oil holding capacity (OHC)
Mixing of hydrogenated Mixing of milk, egg, salt, About 1g of flour sample and 25 ml of commercially olive oil were taken in a 30 vegetable oil and sugar baking powder ml capacity test tube and centrifuged the mixture at 3000 rpm for 20 minutes. The weight of the residue was recorded after decanted the free oil. OHC was calculated as gram of water per gram of dry sample (Rodriguez-Ambriz et al., 2008).
Swelling capacity (SC) After addition of oil mixed together uniformly The swelling capacity of flour sample was evaluated by Okaka and Potter (1977) method.
Emulsion activity (EA) and emulsion stability (ES) Addition of wheat flour, mashed banana and banana The emulsion activity and emulsion stability of sample were evaluated by peel flour Yasumatsu et al., (1972) method.
Flour dispersibility (FD)
Rolling and cutting About 10 g of sample were mixed with distilled water in a cylinder to make a final volume of 100ml, stir vigorously and then allowed it to settle. After 3 hours, settled particles volume was recorded. And finally percentage of flour dispersibility (Kulkarni et al., 1991) was calculated by subtracted it from 100. Baking at 1500 C for 15 min Sensory evaluation of cookies
Figure 1 Flow chart of cookies preparation The sensory evaluation of prepared cookies was carried out by 15 panel members (trained on sensory evaluation) from different food industries in Bangladesh. The Hydrogenated vegetable oil (80g) and sugar (200g) were uniformly mixed panel members evaluated each sample of the cookies for color and appearance, together for 3 minutes at a speed of 10 and 8 rpm by rotary mixture machine. texture & consistency, taste, aroma and overall acceptability. The samples were Then other ingredients such as milk, egg, salt and baking powder (composition rated by 9 point hedonic scale (9-like extremely, 8- like very much, 7-like shows in table-1) were mixed together at a speed of 6 rpm. After mixing above moderately, 6- like slightly, 5-neither like or dislike, 4-dislike, 3-dislike ingredients then added vegetable oil and uniformly mixed for 3 minutes at a moderately, 2-dislike very much, 1-dislike extremely). Score were collected and speed of 8 rpm. Finally wheat flour, mashed banana and banana peel flour were analyzed statistically. The score 6 or more were considered as acceptable for the added and uniformly mixed well together. The batter were rolled out and cut consumer acceptability. 0 uniformly and send it to baking chamber and baked at 150 C temperatures for 15-16 minutes. When the color of cookies turned to light brown color it indicates Proximate analyses of composite flour and cookies the finished baked cookies. After baking, cookies were then cooled and packed in plastic packets. The ready to eat cookies were than stored at room temperature for Proximate analyses such as moisture, ash, fat, protein and crude fiber of the further analysis. sample were analyzed by using AOAC- 2000 method (AOAC, 2000). Energy value of the sample was calculated by using factor method described by by Table 1 Formulation of cookies with banana and banana peel flour (for 1 kg) Chinma and Igyor and the Atwater factor method is [(9xfat) + (4xcarbohydrate) + Normal Sample 1 Sample 2 Sample 3 (4x protein)] (Chinma and Igyor, 2007) Ingredients cookies (0 (5 %) (10 %) (15 %) %) Physical characteristics of cookies Banana (g) 0.00 25.00 50.00 75.00 Banana peel flour Physical property of cookies is an important attributes which plays an important 0.00 25.00 50.00 75.00 (g) role to determine the consumer acceptability. AACC method was used to determine the diameter and thickness of the cookies (AACC, 2000). The Spread Wheat flour (g) 500.00 450.00 400.00 350.00 Factor (SF) of the cookies was determined according to the following formula: Egg (g) 35.00 35.00 35.00 35.00 SF = (Diameter/Thickness × Correction Factor) × 10, where correction factor was 1.0 in this study at constant atmospheric pressure. Hydrogenated 80.00 80.00 80.00 80.00 vegetable oil (g) Statistical analysis Whole milk 80.00 80.00 80.00 80.00 Sugar (g) 200.00 200.00 200.00 200.00 All the sample analyses were performed in triplicate and descriptive statistics were analyzed by using SPSS (Statistical Package for the Social Science) Soybean Oil (g) 100.00 100.00 100.00 100.00 software package version 16.0 (SPSS Inc., Chicago, IL, USA) for all variables. Salt (g) 2.5.00 2.50 2.50 2.50 All of the values are expressed as the mean of three individual replicates ± Standard Error Mean (SEM). All data were subjected statistically to ANOVA test Baking powder (g) 2.5.00 2.50 2.50 2.50 (analysis of variance) and mean value were separated using Tukey’s test (Steele Torrie, 1980). Differences between mean values were considered to be significant at p < 0.05.
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RESULTS AND DISCUSSION highest in banana peel flour (4.47%) on the other hand the lowest content found in the banana. Banana peel flour had a significantly higher amount of ash content Chemical composition of raw material, different functional properties of banana (7.66%) than banana (1.27%) and wheat flour (0.44%). Percentage of ash content peel flour and wheat flour, physical and nutritional properties of cookies were in banana peel flour is relatively higher than those reported in mung bean flour analyzed by using standard procedures. Determination of functional properties 3.42% by Kavitha & Parimalavalli (2014) and 2.6-3.5% in banana flour (Da was important as it show the interaction among the composition, confirmation, Mota et al., 2000). The high crude ash content of banana peel flour can be structure, and physicochemical properties of protein and other food components attributed to relatively high mineral content. Banana peel flour is rich in iron, (Kinsella, 1976). The effect of incorporation of different proportions of banana zinc, calcium, magnesium, phosphorus, and potassium (Michel et al., 2016). peel flour on nutritional and sensorial properties of cookies were also analyzed. Similarly, banana is rich in potassium (Bezemek, 2015). The crude fiber content was also significantly high in banana peel flour (8.82%) than banana (1.34%) and Chemical composition of raw materials wheat flour (0.34%). The high fibers content in banana peel flour (11.81g/100g dry peel) also observed by Romelle et al., (2016). Banana peels contain both The raw materials, i.e., banana, banana peel flour and wheat flour were analyzed soluble and insoluble dietary fiber. Happi Emaga et al., (2007) found insoluble for chemical composition and the data are presented in table 2. Moisture content dietary fiber; cellulose 7 to 12 g/100 g, lignin 6.4 to 9.6 g/100 g and is one of the important indicators which influence shelf life, changes of flavor hemicelluloses 6.4 to 8.4 g/100 g, in banana peels and also found soluble dietary and enzyme activity and storage stability of flours. Research findings suggest that fiber that is pectin 13.0 to 21.7 g/100 g. High fiber diet are very important for flours having more than 14% moisture content or 0.7 to 0.9 aw are prone to mold adult, in that, it promotes the wave-like contraction that helps to move food growth and infestation by insects (Manley, 2000; Ayub, 2003). Banana pulp had through the intestine and also easing the passage of waste and expanding the the highest moisture content (74.77%) whereas banana peel flour contained only inside walls of the colon to make an anti-constipation effect (Eromosele and 9.40% moisture and wheat flour had 10.90% moisture. These results are quite Eromosele, 1993). Crude fibers are known to aid digestion, absorb water and similar to the results found in sweet potato flour by Singh et al., (2008). Moisture make stools larger and softer so as prevent constipation (Ayoola and Adeyeye, content in flour is mainly affected by amount of moisture used for wheat 2009). The high level of fiber in the plantain suggests that it is capable of conditioning and also for type of wheat milling. Another functional component of promoting digestion as fibers are known to aid and speed up the excretion of flour is percentage of protein content by which flour specification, flour condition wastes and toxins from the body, and also prevent colon cancer as it prevents and quality of the product depends. Protein content varies from 6-20% in waste or toxins to stay in the intestine for too long (Ogbonna et al., 2016) . As different varieties of wheat (Kent, 1983). The percentage of protein of the banana carbohydrate content of flours calculated by difference method, it varied from and banana peel flour was found quite low when compared with wheat flour. 19.09 to 77.94%. These values are quite lower than those found in wheat flour 13.20 % by Raihan and Saini (2017) and 13.60% by Juma et al., (2018). Fat percentage was found
Table 2 Chemical composition of raw materials Sample Moisture (%) Ash (%) Protein (%) Fat (%) Fiber (%) Carbohydrate (%) Banana 74.77 ± 1.12 1.27 ± 0.56 3.25 ± 0.16 0.28 ± 0.06 1.34 ± 0.09 19.09 ± 2.47 Banana peel flour 9.40 ± 2.26 7.66 ± 0.84 2.73 ± 0.10 4.47 ± 0.17 8.82 ± 0.17 66.92 ± 3.56 Wheat flour 10.90 ± 2.43 0.44 ± 0.12 8.96 ± 0.18 1.42 ± 0.15 0.34 ± 0.16 77.94 ± 4.12 Data are expressed as Mean ± Standard Error Mean of three replicate measurements
Functional properties of wheat and banana peel flour was 0.63 g/g dry sample than OHC of banana peel flour that was 0.44 g/g dry sample which was also statistically significant (P<0.05). These findings are quite Test of functional properties is very important because it describes how similar to the results observed by Kaushal et al., (2012). Another functional ingredients behave during preparation and cooking. It also describes how they property of flour is swelling capacity and it depends on variety, size of particles affect the quality of finished food product in terms of looks, tastes, and feels. and types of processing methods. A significant (P<0.05) difference was found in WHC, OHC, SC, EA, ES and FD are important properties of flour both of swelling capacity of wheat flour and banana peel flour. Swelling capacity of physicochemical and technological point of views. WHC is the amount of water wheat flour and banana peel flour found 15 ml and 13.50 ml respectively. that can be bound in fiber until the application of external force. In this study, a Emulsion Activity and Emulsion Stability both were found higher in wheat flour significant (P<0.05) difference was found in WHC of banana peel flour and (41.20% and 35.10%) than banana peel flour (36% and 30.50%) respectively wheat flour. Mean WHC of banana peel flour found slightly higher (4.58 g/g dry which was statistically significant (P<0.05). A significant (P<0.05) variation was sample) than WHC of wheat flour (3.19 g/g dry sample) (Table-3). These found in flour dispersibility test of banana peel flour and wheat flour. About 61% findings are quite low compared to dietary fiber found by Larrauri et al., (1996) flour dispersibility found in banana peel flour whereas 66.56% found in whole in mango (12 to 15 g water/g dry sample) and mango peel (11 to 12 g water/g dry wheat flour. sample). Whereas, the mean OHC of wheat flour was found slightly higher that
Table 3 Functional properties of wheat flour and banana peel flour Properties Wheat flour Banana peel flour P value Water holding capacity (g water/g dry sample) 3.19 ± 0.12 4.58 ± 0.08 0.000 Oil holding capacity (g oil/g dry sample) 0.63 ± 0.14 0.44 ± 0.22 0.004 Swelling capacity (ml) 15.00 ± 0.92 13.50 ± 0.85 0.001 Emulsion Activity (%) 41.20 ± 3.42 36.00 ± 2.64 0.000 Emulsion Stability (%) 35.10 ± 2.12 30.50 ± 3.84 0.000 Flour Dispersibility (%) 66.56 ± 1.28 61.00 ± 3.20 0.000 Data are expressed as Mean ± Standard Error Mean of three replicate measurements. Statistically significant p<0.05 when compared wheat flour to banana peel flour.
Nutritional values of the cookies 1996). The substitution of banana and banana peel flour to refined wheat flour at levels of 5–15% showed a lowering effect on the protein contents of the cookies According to Table 4, with increasing the substitution of banana and banana peel (Table 4). A statistically significant variation (P<0.05) was also found in protein flour, the moisture content was increased in cookies. Moisture content ranged content of all sample cookies. Possible reason for this may be higher percentage from 2.28% to 3.35% and highest moisture content were observed in 10% banana of protein in wheat flour than banana and banana peel flour. This finding is quite and banana peel flour cookies. Moisture content increased with the incorporation lower than the cookies made with 100% pigeon pea flour (12.97% protein) of banana and banana peel flour which was statistically significant (P<0.05). The reported by Okpala and Okoli (2011). The protein content found in banana and water binding capacity of banana peel flour found quite higher than the water banana peel flour cookies was close agreement to the cookies of taro-wheat blend binding capacity of wheat flour. These findings are quite similar to the study of (Ojinnaka. 2009). There was no major distinction found in the fat percentage of Bertagnolli et al. (2014), who found that cookies made from composite flour that the cookies made by substitution of 5 to 15% banana and banana peel flour. contain guava peel flour has moisture ranged from 2.7 and 4.9%. Maximum There was no significant variation (P>0.05) found in cookies except sample-2 variation in moisture content was found 2.47 to 8.75% in commercially available and sample-3 cookies. biscuits whether fat content varies between 1.04 to 14.82% (Semwat et al.,
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Table 4 Nutritional values of the cookies Nutrients Normal cookies (0%) Sample 1 (5%) Sample 2 (10%) Sample 3 (15%) Moisture (%) 2.28 ± 0.12a 2.87 ± 0.02b 3.35 ± 0.14c 3.02 ± 0.11d Protein (%) 10.19 ± 1.08a 8.53 ± 1.03b 7.20 ± 1.24c 6.94 ± 0.20d Fat (%) 25.96 ± 1.34a 25.80 ± 0.46a 27.16 ± 0.86a 21.96 ± 0 .67b Ash (%) 1.63 ± 0.04a 2.42 ± 0.08b 2.72 ± 0.02c 3.15 ± 0.53d Crude fiber (%) 0.41 ± 0.02a 0.65 ± 0.03b 0.91 ± 0.04b 1.22 ± 0.10b Carbohydrate (%) 59.53 ± 2.06a 59.73 ± 0.86b 58.66 ± 0.92c 63.71 ± 1.42d Energy (kcal/100g) 513a 505b 508c 480d Data are expressed as Mean ± Standard Error Mean of three replicate measurements. Values in the same row with different superscripts are statistically significant from each other (p < 0.05). Normal cookies=100% wheat flour, Sample-1=Wheat flour substituted with 5% (2.5%) banana + (2.5%) banana peel flour, Sample-2=Wheat flour substituted with 10% (5%) banana + (5%) banana peel flour, Sample-3=Wheat flour substituted with 15% (2.5%) banana + (2.5%) banana peel flour.
A noteworthy difference (P<0.05) was found in ash content of all sample cookies found between normal and samples-1 cookies. The highest amount of dietary and it increased from 1.63 to 3.15%. The maximum ash content (3.15%) was fiber was found in 15% banana and banana peel flour cookies on the other hand, found in 15% banana and banana peel flour cookies followed by 10% (2.72%), the lowest amount found in cookies made by without addition of banana and while the lowest value in normal cookies (1.63%). Increasing trend of ash content banana peel flour. These findings are quite similar to the results obtained by in cookies with increasing substitution of banana and banana peel flour may be Nassar et al., (2008) who also found increasing pattern of fiber with increasing relate to higher ash content in the banana peel flour than wheat flour. Similar substation of citrus by-products flour with wheat flour. Increasing trend of crude Increasing pattern in ash content with increasing pitaya peel flour substitution in fiber content also observed by Inyang and Wayo (2005) in their sesame fortified cookies also observed by Bala et al., (2015) and increasing nature related with cookies from 0.46 to 1.09%. This study fulfilled the recommended range of FAO the higher ash content in the pitaya peel flour (10.57%) than refined flour and WHO (1994), as they suggested not to add more than 5 g dietary fiber per (0.98%). The fiber content of the cookies has increased with increased the 100 g dry matter. substitution of banana and banana peel flour but significant differences (P<0.05)
Table 5 Crude fiber and ash improving effect of banana and banana peel flour Normal cookies (0%) Sample-3 (15%) Difference Nutrient Increased amount (Sample-3- Normal cookies) Crude Ash (%) 1.63 3.15 1.52 93.25% Crude fiber (%) 0.41 1.22 0.81 197.56%
Carbohydrate content of prepared cookies was calculated by difference method Physical characteristics of cookies and there was no remarkable variation found in carbohydrate content of cookies. Carbohydrate content found highest in 15% banana and banana peel flour cookies The cookies were also analyzed for their physical properties such as thickness, followed by 5% banana and banana peel flour cookies. These results are in close diameter and spread factor shown in table 6. A significant variation (P<0.05) relation with the cookies made from cocoyam-wheat blends (Ojinnaka, observed in diameter and thickness of the cookies. The cookies diameter ranged Akobundu and Iwe, 2009). The energy values of the prepared cookies ranged from 5.75 cm to 5.86 cm. while thickness of the cookies ranged from 1.22 cm to from 480 to 513 Kcal /100 g. The slight variation on energy value due to the 1.42 centimeter. The highest thickness was found in cookies made by without variation of protein and fat content of cookies outweighs the decrease in addition of banana and banana peel flour. As spread factor was calculated from carbohydrate content. diameter and thickness of the cookies there was a significant variation (P<0.05) found in spread factor. The highest spread factor was found in 10% banana and banana peel flour cookies and lowest one found in normal cookies. A similar inclination in cookies observed by Chinma and Gernah (2007) made from mango, cassava and soybean composite flour. Higher spread ratios in cookies having are considered the most desirable (Kirssel and Prentice, 1979).
Table 6 Physical characteristics of cookies Physical properties Normal cookies (0%) Sample 1 (5%) Sample 2 (10%) Sample 3 (15%) Diameter (cm) 5.82 ± 0.12a 5.76 ± 0.03b 5.86 ± 0.22c 5.75 ± 0.11d Thickness (cm) 1.42 ± 0.24a 1.30 ± 0.17b 1.22 ± 0.32c 1.28 ± 0.28d Spread Factor 40.99 ± 1.18a 44.31 ± 0.68b 48.03 ± 2.52c 44.92 ± 1.85d Data are expressed as Mean ± Standard Error Mean of three replicate measurements. Values in the same row with different superscripts are statistically significant from each other (p < 0.05). Normal cookies=100% wheat flour, Sample-1=Wheat flour substituted with 5% (2.5%) banana + (2.5%) banana peel flour, Sample-2=Wheat flour substituted with 10% (5%) banana + (5%) banana peel flour, Sample-3=Wheat flour substituted with 15% (2.5%) banana + (2.5%) banana peel flour.
Sensory evaluation of functional cookies wheat flour increased the darkness of cookies may be the prime reason for find objectionable of the cookies. Sensory evaluation of the cookies depends on its appearance, color, flavor and aroma, texture and overall acceptability taste of the sample. All the panelists were Texture and consistency acceptability having excellent capability (trained on sensory evaluation) to evaluate the sensory evaluation of banana and banana peel flour cookies. Texture and Consistency depends upon mainly the rate of development of the dough and the proportion of sugar used. Table 7 shows that the normal cookies Color and appearance acceptability and sample-1 got the highest score on Texture and Consistency of cookies (score 7) and sample-2 got the second highest score on Texture and Consistency. The Table 7 shows the sensory evaluation score of functional cookies supplemented softness of the texture of normal, sample-1 and sample-2 cookies may be the with different level of banana and banana peel flour. Besides normal cookies, main reason for getting the highest score by the panelist. Similarly the hardness sample-2 cookies liked very much by the panelist (score 7.5). On the other hand, of sample-4 than normal cookies may be the reason for getting lowest score. The 15% substitution of banana and banana peel four got the lowest score on color result revealed that, increasing the substitution of banana and banana peel flour and appearance. A statistical significant variation (P<0.05) found on color and increases the hardness of cookies. appearance score. Increasing the supplementation of banana peel flours with
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Taste acceptability taste. About 15% substitution of banana and banana peel flour cookies got the lowest score on taste acceptability taste. Taste is also influenced by the quality of the raw materials used in the processing unit of cookies. No significant variation (P>0.05) found in taste acceptability
Table 7 Comparison of the different organoleptic quality parameters of functional cookies Sample 1 (5%) Sample 2 (10%) Sample 3 (15%) Sensory Evaluation Normal cookies (0%)
Color and Appearance 7.5 ±0.12a 6.1 ± 0.22b 7.5 ± 0.08c 5.6 ± 0.22d Texture and Consistency 7.0 ±0.07a 7.0 ± 0.19a 6.5 ± 0.23a 5.4 ± 0.11a Taste 7.5 ± 0.08a 7.0 ± 0.09a 7.0 ± 0.13a 6.3 ± 0.80a Aroma 7.0 ± 0.16a 7.0 ± 0.24b 6.0 ± 0.07c 5.8 ± 0.33d Overall Acceptability 6.5 ± 0.28a 6.5 ± 0.36b 7.5 ± 0.16c 5.3 ± 0.08d Data are expressed as Mean ± Standard Error Mean (n=15). Values in the same row with different superscripts are statistically significant from each other (p < 0.05). Normal cookies=100% wheat flour, Sample-1=Wheat flour substituted with 5% (2.5%) banana + (2.5%) banana peel flour, Sample- 2=Wheat flour substituted with 10% (5%) banana + (5%) banana peel flour, Sample-3=Wheat flour substituted with 15% (2.5%) banana + (2.5%) banana peel flour.
Aroma acceptability Bala, A., Gul, K. and Riar, C. S. (2015). Functional and sensory properties of cookies prepared from wheat flour supplemented with cassava and water chestnut The aroma of the products depends on the volatile constituents of raw materials. flours. Cogent Food and Agriculture, 1(1), 1019815. Normal Cookies and sample-1 showed the best aroma acceptability taste score https://doi.org/10.1080/23311932.2015.1019815 comparable to other functional cookies. On the other hand, sample-4 neither like B. B. S. (2006). Year Book of Agricultural Statistics of Bangladesh. Bangladesh nor dislike on flavor by the panelist. The score on aroma acceptability taste was Bureau of Statistics, Planning Division. Ministry of Planning Government of the statistically significant (P<0.05). people’s Republic of Bangladesh, 86p. Bertagnolli, S. M. M., Silveira, M. L. R., Fogaça, A. D. O., Umann, L., & Penna, Overall acceptability N. G. (2014). Bioactive compounds and acceptance of cookies made with Guava peel flour. Journal of food science Technology, 34(2), 303–308. Hedonic scale showed that the 10% substitution of banana and banana peel flour http://dx.doi.org/10.1590/fst.2014.0046 got the highest score on overall acceptability taste. The score on overall Bezemek, M. W. (2015). Bananas: rich in fiber, vitamins, and minerals. acceptability taste was highly significant (P<0.05). Retrieved from http://ceimperial.ucanr.edu/files/218815.pdf on 28 Mar 2017. Chinma, C. E. and Gernah, D. I. (2007). Physichochemical and sensory CONCLUSION properties produced of cookies produced from cassava/ soybean/mango composite flours. Journal of Food Technology, 5(3), 256-260. The outcome of the present study reveals that, higher the substitution of banana Chinma, C. E. and Igyor, M. A. (2007). Micronutrients and anti-nutritional and banana peel flour with wheat flour increased the percentage of crude ash and contents of selected tropical vegetable grown in South East, Nigeria. Nig.Food crude fiber content in cookies. There was a significant variation found in protein, Journal, 25(1), 111-116. http://dx.doi.org/10.4314/nifoj.v25i1.33659 fat, carbohydrate and energy content of prepared cookies. In sensory evaluation Da Mota, R.V., Lajolo, F. M., Ciacco, C. & Cordenunsi, B. R. (2000). test, 10% substitution of banana and banana peel flour cookies were more Composition and functional properties of banana flour from different varieties. acceptable with all quality characteristics by the panel member. So substitution of Starch-Stärke, 52(3), 63-68. https://doi.org/10.1002/(SICI)1521- banana and banana peel flour with wheat flour may be more nutritious and cost 379X(200004)52:2/3<63::AID-STAR63>3.0.CO;2-V effective. Eromosele, I. C and Eromosele, C. O. (1993). Studies on chemical composition and physicochemical properties of seeds of some wild plant (Netherland). Plant DECLARATIONS Food for Human Nutrition, 43(3), 251-256. https://doi.org/10.1007/bf01886227 FAO/WHO. (1994). Codex Alimentarius: foods for special dietary uses Conflict of Interest: Authors have declared that no competing interests exist. (including foods for infants and children). 2ndedn. FAO, Rome. Happi Emaga, T., Andrianaivo, R. H., Wathelet, B., Tchango, J. T. and Paquot, Authors' contributions: The authors contributed equally to the concept, M. (2007). Effects of the stage of maturation and varieties on the chemical experiment, analysis of data and writing of manuscript. All authors read and composition of banana and plantain peels. Food Chemistry, 103(2), 590-600. approved the final manuscript. https://doi.org/10.1016/j.foodchem.2006.09.006 Hossain, M. M., Abdulla, F. & Majumder, A. K. (2016). Forecasting of Banana Funding: The authors provided personal resources for funding this paper. Production in Bangladesh. American Journal of Agricultural and Biological Sciences, 11 (2), 93-99. http://doi:10.3844/ajabssp.2016.93.99 Acknowledgements: The authors would like to offer special gratitude and thanks Inyang, U. E. and Wayo, A. U. (2005). Fortification of cookies with dehulled to Department of Food Technology and Nutritional Science, Mawlana Bhashani sesame seed meal. Journal of Tropical Science, 45(3), 103 –105. Science and Technology University, Santosh, Tangail-1902 for the laboratory https://doi.org/10.1002/ts.2 facilities provided during the present study. Juarez-Garcia, E., Agama-Acevedo, E., Sayago-Ayerdi, S. G., Rodriguez- Ambriz, S. L. & Bello-Perez, L. A. (2006). Composition, digestibility and REFFERENCES application in bread making of banana flour. Plant Foods for Human Nutrition, 61(3), 131–137. http://doi:10.1007/s11130-006-0020-x A. O. A. C. (2000). Official methods of food analysis, 15th edn. Association of Juma, N. A. Alviola and Viena, G. Monterde. (2018). Physicochemical and official Analytical Chemists, Washington, D.C. Functional Properties of Wheat (Triticum aestivum) and Selected Local Floursin A. A. C. C. (2000). Approved Methods of American Association of Cereal the Philippines. Philippine Journal of Science, 147 (3), 419-430. Chemists, 10th Ed., American Association of Cereal Chemists, St. Paul. Kaushal, P., Kumar, V. & Sharma, H. K. (2012). Comparative study of Akobundu, E. N. T., Ubbaonu, C. N. and Ndupuh, C. E. (1998). Studies on the physicochemical, functional, anti-nutritional and pasting properties of taro baking potential of non-wheat composite flours. J Food Sci Technol, 25, 211– (Colocasia esculenta), rice (Oryza sativa), pegion pea (Cajanus cajan) flour and 214. https://doi:10.1016/j.jfca.2006.07.005 their blends. LWT-Food Sci Technol, 48(1), 59–68. Aparicio-Saguilan, A., Sayago-Ayerdi, S. G., Vargas-Torres, A., Juscelino, T., https://doi.org/10.1016/j.lwt.2012.02.028 Ascencio-Otero, T. E., & Bello-Perez, L. A. (2007). Slowly digestible cookies Kavitha, S., Parimalavalli, R. (2014). Effect of processing methods on proximate prepared from resistant starch-rich lintnerized banana starch. Journal of Food composition of cereal and legume flours. J Hum. Nutr. Food Sci, 2, 1051-55. Composition and Analysis, 20(3-4), 175–181. Kent, N. L. (1983).Technology of Cereals. 3rd ed. New York: Pergamon Press. https://doi.org/10.1016/j.jfca.2006.07.005 Kinsella, J. E. (1976). Functional properties of protein in food-A survey. Crit Rev Ayoola, P. B and Adeyeye, A. (2009). Proximate Analysis and Nutrient Food Sci Nutr, 5, 219–225. https://doi.org/10.1080/10408397609527208 Evaluation of Some Nigerian Pawpaw Seeds Varieties, Science Focus, 14(4), Kirssel, L. and Prentice, M. (1979). Protein and fiber enrichment of cookie flour 554-558. with brewer’s spent grain. Cereal Chem, 50(4), 261–265. Ayub, M., Wahab, S. and Durrani, Y. (2003). Effect of Water Activity (Aw) Kulkarni, K. D., Kulkarni, D. N. and Ingle, U. M. (1991). Sorghum malt-based Moisture Content and Total Microbial Count on the Overall Quality of Bread. weaning formulations: preparation, functional properties and nutritive value. International Journal of Agriculture & Biology, 5(3), 274-278. Food and Nutrition Bulletin, 13 (4), 322- 327. https://doi.org/10.1177/156482659101300401
333
J Microbiol Biotech Food Sci / Alam et al. 2020/21 : 10 (3) 329-334
Lakshmi, M., Swarnali, D. M., & Usha, R. (2015). Development and analysis of composite flour bread. J Food Sci Technol, 52(7), 4156–4165. https://doi.org/10.1007/s13197-014-1466-8 Larrauri, J. A., Ruperez, P., Borroto, B. & Saura-Calixto, F. (1996). Mango peels as a new tropical fiber: Preparation and characterization. Lebensmittel- Wissenschaftund Technologie, 29(8), 729–733. https://doi.org/10.1006/fstl.1996.0113 Laura, C. O. & Eric, C. O. (2014). Development of cookies made with cocoyam, fermented sorghum and germinated pigeon pea flour blends using response surface methodology. Journal of Food Science Technology, 51(10), 2671–2677. https://doi.org/10.1007/s13197-012-0749-1 Laura, O., Eric, O. & Emelem, U. (2013). Physico-chemical and sensory properties of cookies made from blends of germinated pigeon pea, fermented sorghum, and cocoyam flours. Food Science & Nutrition, 1(1), 8–14. https://doi.org/10.1002/fsn3.2 Manley, D. 2000. Technology of biscuits, crackers and cookies. 2nd ed. p. 85. London: Ellis Horwood Limited. Michel, C. A. P., Maria, F. P. B., Rafaela, C. P., Juciane, A. R. P., Raimundo, V. S. (2016). Chemical composition of unripe banana peels and pulps flours and its effects on blood glucose of rats. Nutrition & Food Science, 46(4), 504-516. https://doi.org/10.1108/nfs-11-2015-0150 Nassar, A. G., AbdEl-Hamied, A. A. and El-Naggar, E. A. (2008). Effect of citrus by-products flour incorporation on chemical, rheological and organoleptic characteristics of biscuits. World Journal of Agricultural Sciences, 4 (5), 612- 616. Noor Aziah, A. A. and Komathi, C. A. (2009). Acceptability attributes of crackers made from different types of composite flour. International Food Research Journal, 16, 479-482. Ogbonna, O. A., Izundu, A. I., Ikeyi, A. P. & Ohia, G. U. (2016). Proximate Compositions of fruits of Three Musa Species at Three Stages of Development. IOSR Journal of Dental and Medical Sciences (IOSR-JDMS), 15( 6), 107-117. https://doi.10.9790/0853-150611107117 Ojinnaka, M. C., Akobundu, E. N. T. and Iwe, M. O. (2009). Cocoyam starch modification effects on functional, sensory and cookies qualities. Pakistan Journal of Nutrition, 8(5), 558-567. https://doi.org/10.3923/pjn.2009.558.567 Okafor, J. N., Ozumba, A. U., & Solomon, H. M. (2002). Production and acceptability of chinchin fortified with oyster mushroom. Nigeria Food Journal, 18, 19–20. Okaka, J. C., Potter, N. N. (1977). Functional and storage properties of cow pea- wheat flour blends in bread making. J Food Sci, 42(3), 828–833. https://doi.org/10.1111/j.1365-2621.1977.tb12614.x Okpala, L. C. and E. C. Okoli. (2011). Nutritional evaluation of cookies produced from blends of pigeon peas, cocoyam and sorghum flour blends. Afr. J. Biotechnol, 10(3), 433–438. Oyeku, O. M., Kupoluyi, C. F., Osibanjo, A. A., Orji, C. U., Ajuebor, F. N., Ajiboshin, I. O. and Asiru W. B. (2008). An economic assessment of commercial production of 10% cassava. J Ind Res Technol, 2(1), 13-20. Raihan, M. and Saini, C. S. (2017). Evaluation of various properties of composite flour from oats, sorghum, amaranth and wheat flour and production of cookies thereof. International Food Research Journal, 24(6), 2278-2284. http://agris.upm.edu.my:8080/dspace/handle/0/16067 Rodriguez-Ambriz, S. L., Islas-Hernández, J. J., Agama-Acevedo, E., Tovar J., & Bello-Pérez, L. A. (2008). Characterization of a fiber-rich powder prepared by liquefaction of unripe banana flour. Food Chemistry, 107(4), 1515–1521. https://doi.org/10.1016/j.foodchem.2007.10.007 Romelle, F. D., Ashwini, R. P. & Ragu, S. M. (2016). Chemical composition of some selected fruit peels. European Journal of Food Science and Technology, 4(4), 12-21. Rungsinee, S. & Natcharee, P. (2007). Oxygen permeability and mechanical properties of banana films. Food Research International, 40(3), 365–370. https://doi.org/10.1016/j.foodres.2006.10.010 Semwat, A. D., Narsimhamurty, M. C. & Arya, S. S. (1996). Composition of some commercially available biscuits. J Food Sci Tech, 33,112–115. Singh, S., Riar, C. S. and Saxena D. C. (2008). Effect of incorporating sweet potato flour to wheat flour on the quality characteristics of cookies. African Journal of Food Science, 2, 65-72. https://doi.org/10.13140/RG.2.2.28963.30249 Steele, R. G. D., & Torrie, J. H. (1980). Principles and Procedures of Statistics. McGraw-Hill Book Company Inc. New York, USA. Yasumatsu, K., Sawada, K., Maritaka, S., Toda, J., Wada, T., Ishi, K. (1972). Whipping and emulsifying properties of soy bean products. Agri Biol Chem, 36(5), 719–727. https://doi.org/10.1080/00021369.1972.10860321
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AMARANTH INFLUENCE ON WHEAT FLOUR DOUGH RHEOLOGY: OPTIMAL PARTICLE SIZE AND AMOUNT OF FLOUR REPLACEMENT
Ionica Coțovanu1, Georgeta Stoenescu2, Silvia Mironeasa*1
Address(es): Mironeasa Silvia, 1 Stefan cel Mare University of Suceava, Faculty of Food Engineering, University Street no.13, 720229, Suceava, Romania. 2Arcada Research laboratory, Arcada Mill, Lunca Siretului, Zona Banda Barbosi 1 bis, 800416, Romania.
*Corresponding author: [email protected] doi: 10.15414/jmbfs.2020.10.3.366-373
ARTICLE INFO ABSTRACT
Received 30. 4. 2020 Knowledge of various amaranth flour (AF) particle sizes influence added in different amounts in refined wheat flour (WF) seems Revised 8. 7. 2020 necessary to properly design systems with expected rheological properties. The study aimed to assess the effects of factors, particle size Accepted 13. 7. 2020 and level of AF added in WF on Falling number index (FN) and dough rheological properties described by Mixolab in a systematic Published 1. 12. 2020 investigation based on design of experiment. The level of AF added in WF significantly influenced the FN values in the linear and quadratic terms (p 0.05), revealing a decrease with the level increase. Particle size played a significant role in increasing water Regular article absorption in linear and quadratic terms (p 0.05), whereas only the linear term has a significant effect (p 0.005) on dough development time and stability. Particle size increase and the interaction between particle size and AF significantly increased torques C2 and C3. AF level increase in WF significantly decreased the torques C4 and C5. The optimization results highlighted that the most suitable combination would be the composite flour with 9.74% AF of 280 μm particle size incorporated in WF. The proposed approach and the information resulted can be relevant for industrial applications, helping to develop products with amaranth with optimum rheological properties, in order to scale-up the production.
Keywords: Rheology, Mixolab, Amaranth flour, Wheat flour, particle size
INTRODUCTION environmental conditions (Akin–Idowu et al., 2017; Venskutonis and Kraujalis, 2013). Amaranth protein presents a high content of essential amino Consumption of alternative crops such as Andean pseudo cereals is gaining acids, especially lysine and methionine (Caselato-Sousa and Amaya-Farfán, popularity among people who adopt a healthy lifestyle. The opportunity to 2012) and thus, it is superior to that of common cereals. The cysteine includes supplement refined wheat flour which has a low level of nutrients with higher sulphur which improved gluten network by inter in sulphur bond.Hence, the nutritional value flour has attracted high interest for manufacturers involved in addition of amaranth flour to wheat flour improves dough characteristics in wheat new bakery products development based on nutrient-dense ingredients. Amaranth flour (Palombini et al., 2013). Comparatively with cereal starch, the amaranth flour is recognized as a functional ingredient due to its high-quality nutritional starch is found in lower amounts and is located in the perisperm, not in the and technological properties, especially in baking processes, and due its multiple endosperm (Inglett et al., 2015; Valcárcel-Yamani and Lannes, 2012). uses for processing and industrialization (Sanz-Penella et al., 2013). Besides the Amaranth starch presents much lower amylose content than other cereal starches, excellent nutritional value, amaranth grain present a potential agronomic whereas amylopectin content is high (Venskutonis and Kraujalis, 2013; importance across the world, being environmentally resistant crop which easily Schoenlechner et al., 2008). Rapidly digestible starch was reported to be about adapted to different conditions (Valcárcel-Yamani and Lannes, 2012; Rosell et 30% d.w. which leads to a lower predicted glycaemic index (87.2) compared to al., 2009). There are more than 60 species of amaranth ,but three species of wheat starch (Kaur et al., 2010). The total fiber content of amaranth seeds varies genus Amaranthus originating in South America are the most important, A. between 11.14 and 20.6%, wich depend on the species and conditions of growing hypochondriacus L. (México), A. cruentus L. (Guatemala) and A. caudatus L. (Venskutonis and Kraujalis, 2013; Valcárcel-Yamani and Lannes, 2012); the (Peru and other Andean countries) being cultivated for grain production soluble fiber being in high quantity (4.2%) (Caselato-Sousa and Amaya- (Cárdenas-Hernández et al., 2016; Kaur et al., 2010). Because amaranth does Farfán, 2012). Other researchers found that the grain amaranth contains 3 times not include gluten, makes it to be a nutritious food ingredient which contain high the amount of fibre in wheat (Dhellot et al., 2006). The lipid content of amaranth level of protein and carbohydrate that require the needs of the people with celiac is higher than cereal grains, ranging from 5.7 to 16.7% (Kraujalis and disease, the individuals allergic to wheat, and the vegetarian persons. Health- Venskutonis, 2013; Alvarez-Jubete et al., 2009). Theatty acids founds in high promoting attributes of amaranth seeds reside in hypocholesterolemic activity, concentration in amaranth oils were palmitic (19%), oleic (26%) and linoleic improves the immune system, antitumor effect, decreases blood glucose levels, (47%) (Caselato-Sousa and Amaya-Farfán, 2012). The amaranth fats are action acts on liver functions, hypertension, antienemiceffect, antioxidantactivity, found to be generally stable against oxidation, due to the presence of high amount celiac disease and antiallergic action (Caselato-Sousa and Amaya-Farfán, of vitamin E. Valcárcel-Yamani and Lannes (2012) reported that amaranth 2012). contains bioactive compunds: riboflavin (0.19-0.23%) and ascorbic acid (4.5%). In addition to being source of carbohydrate and a high quality protein with a well Some Amaranthus species contains other minor components such as phytosterols, balanced amino acid composition, the amaranth seed provides dietary fiber, lipids waxes and terpene alcohols, which primarily depend on plant species and cultivar rich in unsaturated fatty acids, adequate levels of minerals (zinc, copper, and method. In respect to the minerals, calcium, magnesium, iron and zinc can be manganese), vitamins (thiamine, niacin, riboflavin, and folate) and significant found in high amounts in comparison with wheat, the content being influenced by amounts of other bioactive components such as squalene, tocopherols, phenolic environmental conditions (Valcárcel-Yamani and Lannes, 2012; Alvarez- compounds (Chauhan et al., 2015; Inglett et al., 2015; Venskutonis and Jubete et al., 2009). Other bioactive compounds (total phenolic acids) were Kraujalis, 2013) that have known effects on human health (Kalinova and found in amaranth seeds (Repo-Carrasco-Valencia et al., 2010), being a source Dadakova, 2009). The researchers have found varying values of grain amaranth of antioxidant valuable phenolic compounds (de la Rosa et al., 2009). protein composition, from 13.10 to 21.00%, depending on variety and
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Some research has shown an improved nutritional quality of bakery products protein content 17.00% (SR EN ISO 20483:2007), and ash content 3.00% (SR when wheat flour is supplemented with amaranth, depending on the percentage ISO 2171:2009). The amaranth seeds were grounded in a laboratory grinder added. There were studies revealing that amaranth substitution of 10–20% (Grain Mill, KitchenAid, Model 5KGM, Italy) and sieved through a Retsch improve some rheological properties of the dough without damaging quality. Vibratory Sieve Shaker AS 200 basic (Haan, Germany) to obtain amaranth flour Amaranth flour was investigated as a replacement ingredient of wheat flour in (AF) at three different particle size: large, L 300 μm, medium, 180 μm M various baked products such as bread, cakes, biscuits, pasta (Sanz-Penella et al., 300 μm and small fractions, S 180 μm. 2013; Capriles et al., 2008; Vitali et al., 2008; Sudha and Leelavathi, 2012). Most of the studies reported that amaranth has negative effect on loaf volume: Experimental design and statistics this is due mainly to an excess amount addition which reduces gluten content of composite flour, and also leads to disruption of the starch-gluten matrix, thereby The effects of two factors, particle size (PS) at 3 levels (L, M, S) and addition negatively influencing dough rheological properties. The negative effects can be level of AF in wheat flour at five levels (0, 5, 10, 15 and 20%) on the falling diminished by using an adequate proportion of amaranth in wheat flour. number (FN) index, water absorption of amaranth-wheat flour dough and Particle reduction represents an important operation in food processing which is rheological properties using the Mixolab device as responses were investigated widely applied in the grain milling industry to separate the endosperm from the using response surface methodology (RSM) by means of general factorial design. undesirable components of the whole grain (Ahmed et al., 2019). Several studies The experimental design with real and coded factors values which led to 15 revealed that separation of the flour in different fractions vary in physical and experimental formulations (Table 1) was built to allow the estimation of the main chemical properties and thus provided possibility of production of specialty and interaction effects of the factors. flours for different use products (Sakhare et al., 2013). The remarkable effects of particle size on rheological properties of various flour dough with different Table 1 Real and coded values of formulations factors particle size was reported in some previously investigations (Ahmed et al., 2019; Coded values Real values Run Iuga et al., 2019; Mironeasa et al., 2019; Ahmed et al., 2016; Ahmed et al., X1 X2 Particle size (μm) AF (%) 2015; Moreira et al., 2014), highlighting that particle size and composition of 1 -1.000 -1.000 180 0 the particle fractions influenced significantly the rheological properties in various 2 -1.000 0.500 180 15 ways. Some quality attributes of bread, such as volume and hardness are strongly 3 1.000 0.500 380 15 influenced by the particle size of flours added; a higher particle sizes leads to 4 1.000 0.000 380 10 lower hardness and higher volume (Martínez et al., 2014). Large particle size 5 -1.000 -0.500 180 5 can have a negative impact on the bread quality due to the lower hydration and its 6 -0.333 -0.500 280 5 integration in the gluten viscoelastic structure (Mironeasa et al., 2018; Sanz- 7 -0.333 0.000 280 10 Penella et al., 2013). On the other hand, the lower particle sizes showed a higher 8 1.000 1.000 380 0 water holding capacity due to the surface area rising. The important role of water 9 -1.000 0.000 180 10 content in the final product quality as a plasticizer that significantly affect 10 -1.000 1.000 180 20 rheological behavior has been reported (Amjid et al., 2013; Bhattacharya et al., 11 -0.333 1.000 280 20 2006). The incorporation of large particle size rich in fiber and high levels of 12 -0.333 0.500 280 15 amaranth flour can have a detrimental effect on dough rheological and 13 1.000 1.000 380 20 viscoelastic properties, and on bread functional properties, as well as on its 14 -0.333 -1.000 280 0 organoleptic ones. Previous research have been trying to determine an optimal 15 1.000 -0.500 380 5 amount of amaranth flour which could be used for substitution of wheat flour, but no work has been done to examine effects of particle size of amaranth flour. To To describe the effects of two factors, particle size and amount of AF added in our knowledge, no studies have been made yet to investigate into the main effect refined wheat flour on the fifteen responses, a polynomial quadratic regression of particle size and amaranth seed flour level and interaction effect between equation (Eq.1) was applied where Y is the response variable, bo,b1,b2, b11, b22 and b12 particle size and level of amaranth flour added in wheat flour, on -amylase represent the regression coefficients for intercept, linear, quadratic and interaction effects activity using Falling number tests and on dough characteristics using Mixolab. respectively of X1 and X2 independent variables or factors (particle size of amaranth flour The rheological equipment of a newer generation Mixolab, launched in 2004, has and level added in wheat flour) (Iuga and Mironeasa, 2019). gained more and more popularity in assessing dough behaviour during mixing and also during the heating-cooling stage, simulating the behaviour of the dough 2 2 inthe bread making process in only one single test. Mixolab can be used to Y bo b1 X1 b2 X2 b11 X1 b22 X2 b12 X1 X 2 feature the rheological behaviour of dough subjected to a dual mixing and (1) temperature constraints (Moreira et al., 2012), allowing an extremely complex assessment of the changes. The Mixolab has been applied to evaluate the impact The multiple linear regression analysis was applied to fit the data obtained for of particle size on quality attributes of reconstituted whole wheat flour (Wang et each response to linear, two factor interactions, quadratic and cubic models and al., 2016; Liu et al., 2016) as well as the influence of milling whole wheat grains the most adequately predictive model was selected by means of sequential F-test, of different particle size on the thermo-mechanical properties (Bressiani et al., coefficients of determination (R2), adjusted coefficients of determination (Adj.- 2019). R2), and significant probabilities. In order to determine the validity of the model Response surface methodology (RSM) RSM is a useful statistical tools which can for each response, the experimental and predictive values were compared by be employed in order to diminish the number of samples necessary to evaluate paired t test (p 0.05). Analysis of variance (ANOVA) was performed to the factors that affect the process and their interaction implying mathematical evaluate the statistical significance of the coefficients in each predictive model. models and needing small time resources (Mironeasa and Mironeasa, 2019). The polynomial response surfaces were generated showing the combined effect Multiple-response process optimization technique has been successfully applied, of the factors, particle size and addition level. Stat Ease Design-Expert 7.1 in conjunction with desirability function for optimization of different composite software (trial version) was employed to carry out the experimental design, the flour formulation (Mironeasa et al., 2019a; Iuga and Mironeasa, 2019; Marcin model analysis, generation of the response surface plots, test of model adequacy et al., 2016). and the optimum level of formulation factors finding (Mironeasa et al., 2019a). Therefore, the aim of this study was to assess the effects of different amaranth The multiple responses analysis was applied to the predictive models fitted in flour particle size additions at different levels in refined wheat flour on dough order to find the optimal value of the factors, amaranth flour particle size and rheological properties and to determine the optimal formulation of amaranth addition level. In the optimization process, each predicted response is particle size-level added which will give the adequate dough rheological transformed into an individual desirability function, dn which comprise the properties. desired and researcher’s priorities when building the optimization procedure for each of the factors. The individual desirability functions are then combined into a MATERIAL AND METHODS single composite response, named overall desirability function, D (Eq. 2)
computed as the geometric mean of the individual desirability dn which varies Basic materials from 0 to 1 (Wu et al., 2011).
Commercial refined wheat flour (WF) of 650 type used in this study was (1/n) acquiered from the local market from Romania (MOPAN,Suceava) and consisted D = (d1d2 … dn) (2) of 14.00% moisture (ICC 110/1), 12.60% protein (ICC 105/2), 1.40% fat (ICC 136), 0.65% ash (ICC 104/1), 30.00% wet gluten (ICC 106/1), 6.00 mm gluten where d1,d2,….dn represent the individual desirability function and n is the deformation index (SR 90:2007), and 312 s falling number index (ICC 107/1), number of responses. being a good flour for bread making according to the Romanian standard SR 877:1996. The amaranth grains, acquired from S.C. SOLARIS PLANT S.R.L. For the numerical optimization, dough stability (ST) and starch gelatinization, as (Ilfov, Romania), presented the following analytical characteristics: moisture the differences between C3 and C2 (C3-2) were set as maximum, while protein content 14% (SR EN ISO 665:2003), fat content 8.00% (SR EN ISO 659:2009), weakening, as the differences between C1 and C2 (C1-2) and starch
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recrystallization during paste cooling, as the differences between C5 and C4 (C5- C3 torques (N·m) which measures the starch gelatinization, the difference 4) were set as minimum. The level of all remaining responses which considered between the C3 and C4 torques (N·m) which is related to starch breakdown and in this study was kept within range. the difference between the C5 and C4 torques (N·m) related to retrogradation of starch during paste cooling were recorded. Falling Number Evaluation RESULTS AND DISCUSSION The Falling number (FN) method, based on viscosity, was applied in order to achieve a proximate estimation evaluation of the -amylase activity in the The results obtained are presented in Table 2 and shows the effects of composite flour formulated. The FN values were determined according to ICC formulation factors, particle size and addition level on FN value of amaranth- standard method 107/1. wheat composite flour and Mixolab rheological parameters presented as their corresponding regression coefficients in the quadratic model. The accuracy test Evaluation of rheological characteristics by Mixolab of the model (Table 2) showed that the quadratic model predict properly the
Mixing and starch pasting characteristics of samples formulation according to the studied parameters as a function of the formulation factor. experimental design (Table 1) were studied using the Mixolab equipment The falling number (FN) index values of amaranth-wheat composite flours (Chopin, Tripette et Renaud, Paris, France) with standard option “Copin+” formulated ranged from 289 to 321 s. As shown in Table 2, the level of protocol. The composite flour rheological characteristics were determined amaranth flour (AF) added, has a significant negative effect on the FN, while the according to ICC standard method no. 173 (ICC, 2010). The protocol used for particle size has a non-significant effect (p 0.05). FN was significantly determining the rheological behaviour of amaranth-wheat flour was: dough correlated (p 0.05) with the interaction effect of AF addition level in WF and mixing at constant mixing rate of 80 rpm and temperature of 30C during 8 min, particle size and, also with quadratic effect of AF addition level. By ANOVA, the 2 heating rate 4C/min up to 90C, cooling rate of 4C/min up to 50C, and total correlation coefficient value (R ) of this model was determined to be 0.70, while 2 time to run the analysis of 45 min. All the samples were made at the optimum the adjusted determination coefficient value (Adj.-R ) was 0.55, showing that the hydration level of the composite flour formulated that lets to achieve the regression model defined well the real -amylase activity of the composite flour optimum consistency of dough corresponding to the C1 torque value of 1.1 N·m. through FN index. The negative coefficient of the AF addition level indicated Several preliminary mixing tests were made in order to determine the optimum that the FN index of amaranth-wheat composite flour decreased with increasing hydration level to reach the maximum consistency of dough (C1 = 1.1 N·m) and levels. The effects of particle sizes and AF addition level is shown in Figure 1a, then complete test has been applied to determine the dough properties as function indicating a decrease of FN index with AF addition increase. This trend suggests of mixing and temperature. The following mixing parameters from the registered an increase of the -amylase activity in the composite flour when the AF addition Mixolab curves were determined: water absorption, WA (%) or percentage of level in WF increases, knowing that the FN index is inversely correlated with - water needed to achieve the maximum consistency torque C1 of 1.1 N·m, dough amylase activity in flour (Struyf et al., 2016).The increase of -amylase activity development time, DDT (min) - the time to reach the maximum torque at 30C of the mixture is related to the metalloenzyme character of α-amylase whose and stability, ST (min) as time in min until the loss of consistency does not activity depends on the presence of calcium ions in its structure (Suandarram et exceeds 11% of the C1 torque. When dough temperature increases, due to the al., 2014). It is known that amaranth seed contain a high level of calcium (Sanz- protein reduction the minimum consistency, C2 (N·m) torque was recorded. Penella et al., 2013; Alvarez-Jubete et al., 2009) and therefore an increased Then, when dough temperature increases to certain values, the starch level of amaranth flour in the amaranth-wheat composite flour will result in an gelatinization was given through the values of maximum consistency, C3 torque increase of -amylase activity mixture, improving final product quality. (N·m), the stability of starch gel by minimum consistency, C4 torque (N·m) and starch retrogradation when cooling dough through maximum consistency, C5 torque (N·m). In addition, the difference between the C1 and C2 torques (N·m) which is a measure of the protein weakening, the difference between the C2 and
Table 2 Regression coefficients in the predictive models represented as a function of variations in the formulation factors for FN value and wheat flour dough Mixolab characteristics Parameters Factorsb FN WA DT ST C2 C1-2 C3 C3-2 C4 C3-4 C5 C5-4 (s) (%) (min) (min) (Nm) (Nm) (Nm) (Nm) (Nm) (Nm) (Nm) (Nm) Constant 316.25 58.450 2.890 9.950 0.520 0.600 1.760 1.250 1.580 0.180 2.380 0.800 A ns -0.780*** ns 0.840*** 0.045*** -0.050*** 0.099*** 0.054*** ns 0.073*** ns ns B -4.730** 0.170* 1.670*** -0.830** -0.036*** 0.029*** -0.180*** -0.150*** -0.330*** 0.015*** -0.620*** -0.260*** A x B -5.850** -0.770*** ns 0.780** 0.042*** -0.044*** 0.081*** 0.040** ns 0.077*** ns ns A2 ns -0.420** ns ns -0.035*** ns ns ns ns ns ns ns B2 -7.900** 0.580** ns ns ns 0.032** ns ns ns ns ns ns
R2 0.70 0.93 0.72 0.83 0.91 0.93 0.95 0.95 0.95 0.96 0.93 0.88 Adj.-R2 0.55 0.89 0.57 0.73 0.87 0.89 0.93 0.92 0.92 0.94 0.89 0.82 p-value 0.0205 < 0.0001 < 0.0202 0.0028 < 0.0001 < 0.0001 < 0.0001 < 0.0001 < 0.0001 < 0.0001 < 0.0001 0.0005 *** p < 0.005, ** p < 0.05, * p < 0.5, A –Particle size (m), B – level of amaranth flour added in refined wheat flour (%), R2,Adj.-R2 – measures of model fit, FN – Falling number, WA – water absorption, DT – development time, ST – stability, C2 – minimum consistency during protein weakening stage, C1-2 – difference between torques C1 and C2, C3 – maximum consistency during starch gelatinization stage, C3-2 – difference between torques C3 and C2, C4 – minimum consistency corresponding to hot starch stability gel, C3-4 – difference between torques C3 and C4, C5 – maximum consistency during starch retrogradation stage, C5-4 – difference between torques C5 and C4.
The addition of amaranth flour may improve the quality of the final baked Effects of formulation samples on dough rheological properties during products prepared especially from flour with a low -amylase activity as it is mixing stage known that -amylase provides fermentable sugars leading to increased bread volume, improves crumb structure, intensifies crust colour (Mironeasa and During initial Mixolab kneading stage, the formation of a three-dimensional Codină, 2017; Struyf et al., 2016). viscoelastic structure with gas-retaining properties is related to the distribution of the material, the disruption of the initially spherical protein particles and the Effects of the amaranth flour particle size and the levels added in refined hydration of the flour compounds that occur together with the stretching and wheat flour on the Mixolab rheological parameters alignment of the proteins (Rosell et al., 2007; Angioloni and Collar, 2009). When refined wheat flour was replaced by AF, dough consistency and stability Amaranth flour addition of different particle sizes at various levels in refined that are mainly related to the aggregates formation as a result of hydrogen wheat flour has deeply changed the rheological behaviour of dough measured bonding or proteins linking through disulfide or dityrosine bonds, undergo with Mixolab, device which provides information on the protein quality and modifies. AF addition in WF changes dough consistency at a given amount of strength, amylolytic activity and starch gelling (Codină and Mironeasa, 2013). added water depending on AF particle size and level. It is well-known that water Parameters registered in five different stages of the Mixolab curve describe absorption capacity influences the baking process, being also a significant dough rheological properties during mixing, measures the speed of weakening of economic indicator that affects the volume yield of the finite product. Water protein due to mechanical work and heat, starch gelatinization, the stability of absorption (WA) of formulated samples varies from 56.70 to 60.50%. The effect hot-formed starch paste and starch retrogradation during the cooling stage. of particle size and AF addition level on WA of amaranth-wheat composite flour dough as their corresponding regression coefficients in the quadratic model
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indicates is shown in Table 2. The model showed that the linear and quadratic terms of particle size has a significant negative influence on the WA suggesting that with particle size decrease WA increases due to the greater surface area of the particles which absorbed more water These results are in agreement with the observation reported in the literature by Ahmed et al. (2019), Drakos et al. (2017) for quinoa, rye and barley flours. The linear term of the AF addition level in WF has a non-significant effect (p 0.05), but the quadratic term showed a significant effect (p 0.05) on WA. Also, WA was significantly correlated with the interaction effect of particle size and AF addition level and, also with quadratic effect of particle size (Table 2). The test for the precision of the model indicates that the quadratic model can be suitable to predict WA as a function of the formulation factors. The R2 and Adj.-R2 values showed that the regression model fitted to the experimental data of WA parameters showed higher R2 value (0.93). The effect of particle size and AF level added in WF on the WA of amaranth-wheat composite flour dough is shown in Figure 1b. The response surface plot indicates that with the interaction effect between particle size and AF level increase, the WA decreased, result which can be expected. This decrease may be explained by the gluten dilution which requires less hydration and (b) therefore a lower amount of water needed (Mohammed et al., 2014). On the other hand, an increase of WA with particle size decrease was found and can be related to the specific surface area of amaranth starch which is larger than wheat starches and small particle absorb more water compared to large and medium particle fractions. Similar result was reported by Iuga et al. (2019), Mironeasa et al. (2019), Ahmed et al. (2016) whereby small particle size of non-gluten flour increases the values of WA. Amaranth flour (AF) addition influenced the gluten network and changed dough development time (DT). The particle size did not significantly (p 0.05) influence DT, while AF level added in WF has a significant effect (p 0.05) on DT of amaranth-wheat composite flour which ranged from 1.33 to 5.75 min. As shown in Table 2, the quadratic model was found to represent adequately the results for DT, as a function of the formulation factors, the R2 and Adj.-R2 values confirming the adequacy of the model. Figure 1c represents a response surface plot showing the effect of particle size and AF level added in WF on DT, revealing that DT increased abruptly (about 2 - 3 times) as the AF level increased above 10% comparatively to the control, depending on particle size. DT increases considerably in the composite flour with large particle size, showing that a long (c) time is needed between the addition of water and the time when the dough reached the optimal elastic and viscous characteristics. The increase in DT can be explained by the addition of non-gluten flour which affects gluten quality. The DT value is a measure of dough strength and the higher it is the stronger is the dough. Dough stability (ST) was affected (p = 0.02) by the particle size and AF addition level in WF. Regression model for ST (Table 2) showed a significant effect in linear term of particle size, AF addition level, and of the interaction terms between particle size and AF addition level, while the quadratic terms of factors formulation were non-significant (p 0.05). Particle size affects positively ST, while AF addition level negatively influences it. The significant negative effect on dough stability given by the AF addition level may be probably due to gluten dilution because wheat flour was replaced with non-gluten flour and can reflect an increased degree of softening. On the other hand, an increase of dough stability with particle size increase, indicates an increase in the gluten network which denotes the fact that the composite dough with large particle size is able to sustain for a longer time the mechanical treatment during the bread making (d) process. This increase can be related to some components from the particle size fractions because it was shown that amaranth albumins are able to interact with Figure 1 Response surface plot showing the combined effects of amaranth flour gluten proteins through disulfide bonds (Oszvald et al., 2009), increasing dough particle size and levels added in refined wheat flour on: Falling number index (a) ST. The results of ANOVA show that the equations can be suitable to predict and Mixolab parameters during dough development stage: (b) water absorption dough ST as a function of the particle size and AF level incorporated in WF. The (WA), (c) development time (DT) and (d) stability (ST) 2 2 R and Adj.-R values were adequate to confirm the significance of the model. A response surface plot for the stability (Figure 1d) showed that dough ST This behaviour was probably related to the addition of non-gluten flour which significantly decreased with an increase of AF level. diminishes dough viscoelastic properties (Mohamed et al., 2014). The weakening of the gluten network can be probably attributed to an intense incompatibility between the protein spectrum of amaranth and the wheat gluten protein. A similar observation on the incompatibility between the protein spectrum of pulses and the semolina gluten protein was reported by Mohamed et al. (2014).
Effects of formulation samples on protein weakening stage
Due to the mechanical shear and temperature constraint, protein weakening occurs and C2 torque decreased to the minimum value. The value of C2 torque was significantly influenced (p< 0.0001) by the particle size and level of AF addition in WF (Table 2). Through ANOVA, the quadratic model was found to fit adequately the experimental data for C2, having a high coefficient of determination (R2 = 0.91). The particle size and AF addition level and the interaction term between these factors have significant effect on C2. The results showed an increase of C2 as the particle size and the interaction between particle size and AF addition increased (Table 2). A negative effect on C2 torque was given by the linear regression coefficient, suggesting that the increased level of (a) AF added in WF induced a C2 torque decrease (Figure 2a).The decrease of C2
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with the AF level increase can be related to lower water availability in the dough increase and higher level of AF (20%) addition in wheat flour lead to a dropped system due the fact that the water absorption capacity of dough decrease when value for C3 torque (1.32 N·m) which was obtained for small particle size. This particle size and AF addition level increase. C2 torque reduction with particle can be due to the amylase activity of amaranth flour and the amylose-lipid size decrease, especially in the case of small particle size, may be linked either to complexes formed during heating of starch slurries. These results are in a release of water molecules in dough or to the presence of secondary activities. agreement with those found by Lorenz (1981), who reported a gradually decreased of maximum viscosity with amaranth level added in wheat flour increase from 3 to 15%. The decrease of particle size leads to a decrease of C3 torque, the small particles fraction considered as starch-enriched particle cause a lower C3 torque compared to coarse particles that can be considered as fiber- enriched particle fractions. Due to the large surface, the small particle sizes compete for water with starch granules from wheat flour, resulting in a decrease of dough viscosity. Similar observation was made by Ahmed et al. (2019) and Singh et al. (2019) for quinoa and corn flour and they reasoned that the larger particles have smaller surface area with lower swelling, which contributed to higher pasting viscosities. The difference among particle fractions could be related to the microstructure of the amaranth-starch that governs the water absorption and swelling, being also influenced by the composition of the particles. The gelatinization of starch measured by the torque C3 reflects breaking of hydrogen bonds in the crystalline regions and assimilation of water by hydrogen bonds and water absorption by non-starch polysaccharides and protein (Yu et al., 2012).
(a)
(a)
(b) Figure 2 Response surface plot showing the combined effects of amaranth flour particle size and level added in refined wheat flour on Mixolab parameters during protein weakening stage: (a) C2 torque (C2) and (b) difference between torques C1 and C2 (C1-2)
The difference between the points C1 and C2 torque (C1-2) which depicts the protein network strength under increasing heating is significantly influenced by the particle size, AF addition level and the interaction between them, as revealed by the ANOVA (Table 2). Value of C1-2, shows a high coefficient of determination (R2 = 0.93) and was obtained for the quadratic model, statistically significant at p < 0.0001, revealing that this model is fitting well for this data. Major negative effects on C1-2 were given by particle size and by the interaction between particle size and AF addition level. The results showed an increase of C1-2 as the AF value increased, indicating a decrease of protein weakening speed under the effect of temperature increase. With particle size increase, C1-2 decreases. This fact can be due to the changes in protein network structure; (b) proteins become less compact, favouring the enzymatic attacking points and Figure 3 Response surface plot showing the combined effects of amaranth flour leading to a rise in the speed of protein weakening due to the heat. This result can particle size and level added in refined wheat flour on Mixolab parameters during be also related to the area specific surface of amaranth starch that is larger than starch gelatinization stage: (a) C3 torque (C3) and (b) difference between torques for wheat starches and it makes them more sensitive to hydrolysis by α-amylase C3 and C2 (C3-2) than wheat starch (Tester et al., 2004). The effect of particle size and AF addition level in wheat flour on protein network strength under increasing heating The effects of particle size and AF addition level in wheat flour on the difference is shown in Figure 2b, revealing that under combined effect between particle size between C3 and C2 torques (C3-2) expressed as their corresponding regression and AF addition level, the protein network becomes weaker due to heat. coefficients in the quadratic regression model are shown in Table 2. The linear term of particle size indicates a significant positive influence on C3-2, while the Effects of formulation samples on starch gelatinization stage linear term of the AF addition level has a significant negative effect. C3-2 was significantly correlated (p < 0.05) with the interaction effect of particle size and As heating proceeds, the starch changes occurred being highlighted by starch AF addition level in wheat flour. The quadratic regression model represent gelatinization, C3 torque and difference between C3 and C2 torques (C3-2). The adequately the experimental data of C3-2, the R2 value (0.95), confirming the particle size, AF addition level and the interaction between these ones adequacy of the model. The effect of particle size and AF addition level in wheat significantly (p < 0.05) affect peak torque (C3) which is associated with the flour can be seen in Figure 3b, indicating an increase of C3-2 with particle size starch gelatinization process that occur when dough is heated above 60 C. Low increase. This can be probably due to the combined effect of the -amylase amylose content in the amaranth starch (Corke et al., 2015) is the main reason activity increase in composite flour and to the compounds that exist in particle for a significantly lower C3 torque in amaranth-wheat composite flour compared fractions composition of amaranth flour. The hydrolysis process of starch, to the control. Similar observations regarding starch gelatinization process were intensified by a high -amylase activity, will lead to a decrease in viscosity (de reported by Ahmed et al. (2019) for quinoa flour. The linear coefficient of the Souza and Magalhães, 2010) and increase the free water amount from the AF addition level indicates a significant negative influence on the torque C3, dough; however the hydrocolloid molecules from the amaranth flour will while particle size has a positive effect. Through ANOVA, the quadratic model reducethe availability of water for granule swelling (Mironeasa and Codină, was found to fit adequately the experimental result for C3 torque (Table 2). 2017; Pahwa, 2016). According to Figure 3a, C3 gradually decreased with the amaranth substitution
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Effects of formulation samples on the stability of hot-formed starch gel Effects of formulation samples on starch retrogradation
The effects of formulation factors, particle size and AF added amount in wheat The starch retrogradation during the cooling period of the Mixolab is represented flour on the C4 torque corresponding to hot starch stability gel (cooking stability) by the C5 torque and the difference between C5 and C4 peaks (C5-4), showed a significant (p < 0.05) decrease with the AF addition level increase, determinated by starch proportions of amylose and amylopectin (BeMiller, while particle size had a non-significant effect (p 0.05) (Table 2). Data obtained 2019), since amylose recrystallizes faster than amylopectin. Quadratic model was for the quadratic regression model indicates adequately results of C4, indicating a fitted for C5 torque and ANOVA results shows that it is well adjusted to the high R2 value of 0.95. As can be seen in Figure 4a, as level the of AF increased experimental data obtained, being highly significant (R2 = 0.93). The significant and particle size decreased, while C4 torque and the stability of the gel formed effect (p < 0.05) of AF level added in wheat flour was observed on C5 torque, decreased. Paste heating at high temperature results in viscosity drop attributed to indicating low retrogradation and recrystalization, whereas the particle size has a the paste resistance to disintegrate at high temperature. The lowest C4 torque non-significant effect (p 0.05). The low starch retrogradation in amaranth- value (1.09 N·m) was obtained for small particle size at higher level of AF (20%) wheat composite flour may be due to the different nature of the starch, and to the used in wheat flour, whereas the highest value (1.783 N·m) was obtained for small granules of amaranth which present a low tendency toward retrogradation. large particle size at lower level of AF (5%). The decrease of C4 can be due to In addition, the higher levels of phenolics can contribute to retrogradation the particle fraction composition and probably to the compounds present in small lowering (Wu et al., 2009). The structure reorganization of starch may be particle size content, like soluble fiber which can bind water by the hydrogen beneficial for prolonging the hardening of bakery products during storage (Cai et bonds, causing a decrease of available water for the starch granules. Pectic al., 2014). Response surface plot for C5 torque showed that an increased level of polysaccharides and xyloglucans in varying amounts, depending on the fiber AF added in WF decreased C5 torque (Figure 5a). fraction, were found mainly in the amaranth fiber (Wefers et al., 2015). The low amylose content of small particle size can lead to a more fluid gel formation without such a defined three-dimensional structure (Tang and Copeland, 2007), contributing to a decrease in the C4 value.
(a)
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(b) Figure 5 Response surface plot showing the combined effects of amaranth flour particle size and level added in refined wheat flour on Mixolab parameters during starch retrogradation: (a) C5 torque (C5) and (b) difference between torques C5 (b) and C4 (C5-4) Figure 4 Response surface plot showing the combined effects of amaranth flour particle size and level added in refined wheat flour on Mixolab parameters during The quadratic regression model obtained for the difference between C5 and C4 cooking stage: (a) C4 torque (C4) and (b) difference between torques C3 and C4 torques (C5-4) showed a good correlation coefficient (R2 = 0.95). It can be seen (C3-4) from Table 2 that C5-4 was significantly (p < 0.05) influenced by the ASF level added in wheat flour, particle size showing a non-significant effect (p 0.05) on The quadratic regression model was fitted for the difference between points C3 C5-4. The effect of PS and ASF level addition in wheat flour on C5-4 is shown in and C4 torques peak values (C3-4), indicating that the model explains 96% of the Figure 5b. The response surface plot revealing reveals that the lowest value for observed results fluctuation. C3-4 was significantly associated with the C5-4 (0.471 N·m) is obtained at maximum ASF level (20%) of small particle size formulation factors, particle size and AF addition level in WF and with the added in wheat flour. It seems that amaranth flour addition limits starch interaction between them (Table 2). The effect of formulation factors on C3-4 is retrogradation preserving bread freshness. This observation is in agreement with showed in Figure 4b, indicating the ability of the particle size and AF level and of the finding reported by Collar and Angioloni (2014) which shows that amaranth their interaction to increase the C3-4. Since the amaranth flour did not bring flour has a positive impact on bread keeping behaviour during aging, revealing amylase in dough system, as it can be seen from the FN index values variation, long term storage. these parameter values increase as compared to the sample where no amaranth flour is added. All the C3-4 presented high values in the samples with amaranth Optimization flour addition, depending on the particle size. On the other hand, C3-4 increase can suggest reduced gel stability when hot and this may be related to the starch The optimization highlighted that the most suitable composite flour would be damage process. with 9.74% amaranth flour of 280 μm particle size, with predicted responses shown in Table 3. There are no significant differences between the results of the water absorption, dough stability and protein weakening (C2 and C1-2) of optimized and control sample. Therefore, optimal dough formulation can retain
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gas, especially during the early stage of bread similar to the control sample. An BHATTACHARYA, S., NARASIMHA, H.V., &BHATTACHARYA, S. (2006). increase of dough development time can suggest an increase of gluten network Rheology of corn dough with gum arabic: Stress relaxation and two-cycle strength which revealed that the optimal dough can sustain for a longer the compression testing and their relationship with sensory attributes. Journal of mechanical treatment during the bread making process compared to the control. Food Engineering, 74(1), 89-95. The slight decrease of specific parameters in heating stage indicate that the BeMILLER, J.N. (2019). Starches: Molecular and granular structures and optimal formulation can be suitable for bread-making, whereas the parameters in properties: Chemistry for Food Scientists, 3 rd ed. 159–189. Elsevier. the cooling stage explain that products of amaranth flour dough do not present BRESSIANI, J., ORO, T., DA SILVA, P., MONTENEGRO, F., BERTOLIN, T., problems of staling and can be maintained for longer time. GUTKOSKI, L., & GULARTE, M. (2019). Influence of milling whole wheat grains and particle size on thermo-mechanical properties of flour using Mixolab. Table 3 Wheat flour dough control sample and optimized amaranth-wheat flour Czech Journal of Food Sciences, 37(4), 276-284. sample https://doi.org/10.17221/239/2018-CJFS Values CAPRILES, V.D., ALMEIDA, E.L., FERREIRA, R.E., ARÊAS, J.A.G., STEEL, Parameters Optimized sample Control sample C.J., &CHANG, Y.K. (2008). Physical and sensory properties of regular and FN (s) 316.37 313.13 reduced‐fat pound cakes with added amaranth flour. Cereal Chemistry,85, 614– CH (g/100g) 58.45 58.43 8. https://doi.org/10.1094/CCHEM-85-5-0614 DT (min) 2.85 1.43 CÁRDENAS-HERNÁNDEZ, A., BETA, T., LOARCA-PIÑA, G., CASTAÑO- ST (min) 9.97 9.78 TOSTADO, E., NIETO-BARRERA, J. O., &MENDOZA, S. (2016). Improved C2 (Nm) 0.52 0.51 functional properties of pasta: Enrichment with amaranth seed flour and dried C1-2 (Nm) 0.60 0.60 amaranth leaves. Journal of Cereal Science, 72, 84-90. C3 (Nm) 1.77 1.92 https://doi.org/10.1016/j.jcs.2016.09.014 C3-2 (Nm) 1.25 1.41 CASELATO-SOUSA, V.M., & AMAYA-FARFÁN, J. (2012). State of knowledge on amaranth grain: a comprehensive review. Journal of Food Science, C4 (Nm) 1.59 1.89 77(4): 93–104. DOI: 10.1111/j.1750-3841.2012.02645.x C3-4 (Nm) 0.18 0.03 CHAUHAN, A., SAXENA, D. C., & SINGH, S. (2015). Total dietary fibre and 2.40 3.02 C5 (Nm) antioxidant activity of gluten free cookies made from raw and germinated C5-4 (Nm) 0.81 1.13 amaranth (Amaranthus spp.) flour. LWT-Food Science and Technology 63, 939– 945. https://doi.org/10.1016/j.lwt.2015.03.115 The improvement of refined wheat flour with amaranth lead to C3, C4 and C5 COLLAR, C., &ANGIOLONI, A. (2014). Pseudocereals and teff in complex torques decrease due to the effect of wheat starch dilution and also, as a result of breadmaking matrices: impact on lipid dynamics. Journal of Cereal Science, proteins interactions. The origin of the proteins has been suggested to be 59(2), 145–154. https://doi.org/10.1016/j.jcs.2013.12.008 responsible for modifying pasting and gelling properties of cereal starches CORKE, H., CAI, Y.Z., WU, H.X. (2016). Amaranth: overview In: Wrigley C, (Marco and Rosell, 2008), due to the thermal induced gelation process of the Corke H, Seetharaman K, Faubion J, editors. Encyclopedia of food grains. proteins after the aggregation of the polymer chains that cause huge Oxford: Academic Press. p 287–96. modifications in the rheological behaviour of the proteins. However, the starch CAI, L., CHOI, I., HYUN, J.N., JEONG, Y.K., &BAIK, B.K. (2014). Influence retrogradation indicates that bread freshness can be maintained for longer time of bran particle size on bread-baking quality of whole grain wheat flour and compared to the control bread. starch retrogradation. Cereal Chemistry, 91, 65– 71. https://doi.org/10.1094/CCHEM-02-13-0026-R CONCLUSIONS CODINA, G.G., & MIRONEASA, S. (2013). Influence of mixing speed on dough microstructure and rheology. Food Technology and Biotechnology, 51(4), A multi-response optimization study helped us to find the optimal particle size 509-519. https://hrcak.srce.hr/114467 and amaranth level added in refined wheat flour in order to achieved a dough DE LA ROSA, A.B., FOMSGAARD, I.S., LAURSEN, B., MORTENSEN, A.G., with the best rheological characteristics. The effect of particle size and amaranth OLVERA-MARTÍNEZ, L., SILVA-SÁNCHEZ, C., & DE LEÓN- level added on the response variables could be accurately predicted by the RODRÍGUEZ, A. (2009). Amaranth (Amaranthus hypochondriacus) as an quadratic type models. By applying numerical technique and desirability alternative crop for sustainable food production: Phenolic acids and flavonoids function, the optimum particle size and level of amaranth flour addition was with potential impact on its nutraceutical quality. Journal of Cereal Science, obtained. The best formulation giving the desirable rheological characteristics 49(1), 117-121. https://doi.org/10.1016/j.jcs.2008.07.012 comprises of 9.74% amaranth flour of 280 μm particle size. DE SOUZA, P.M., & DE OLIVEIRA e MAGALHÃES, P. (2010). Application of microbial -amylase in industry - A review. Brazilian Journal of Acknowledgments: The authors thank Arcada Research laboratory, Arcada Microbiology, 41(4), 850–861.https://doi.org/10.1590/S1517- Mill, Romania for technical support. 83822010000400004 DHELLOT, J.R., MATOUBA, E., MALOUMBI, M.G., NZIKOU, J.M., REFERENCES NGOMA, D.G.S., LINDER, M., DESOBRY, S., & PARMENTIER, M. (2006). Extraction, chemical composition and nutrional characterization of vegetable AHMED, J., THOMAS, L., & ARFAT, Y.A. (2019). Functional, rheological, oils: Case of Amaranthus hybridus (var 1 and 2) of Congo Brazzaville. African microstructural and antioxidant properties of quinoa flour in dispersions as Journal of Biotechnology, 5(11), 1095–1101. influenced by particle size. Food Research International, 116, 302-311. DRAKOS, A., KYRIAKAKIS, G., EVAGELIOU, V., PROTONOTARIOU, S., https://doi.org/10.1016/j.foodres.2018.08.039 MANDALA, I., & RITZOULIS, C. (2017). Influence of jet milling and particle AHMED, J., AL-ATTAR, H., &ARFAT, Y.A. (2016). Effect of particle size on size on the composition, physicochemical and mechanical properties of barley compositional, functional, pasting and rheological properties of commercial and rye flours. Food Chemistry, 215, 326-332. water chestnut flour. Food Hydrocolloids, 52, 888-895. https://doi.org/10.1016/j.foodchem.2016.07.169 https://doi.org/10.1016/j.foodhyd.2015.08.028 INGLETT, G.E., CHEN, D., & LIU, S.X. (2015). Physical properties of gluten- AHMED, J., AL-JASSAR, S., &THOMAS, L. (2015). A comparison in free sugar cookies made from amaranth–oat composites. LWT-Food Science and rheological, thermal, and structural properties between Indian Basmati and Technology, 63, 214– 220. https://doi.org/10.1016/j.lwt.2015.03.056 Egyptian Giza rice flour dispersions as influenced by particle size. Food ICC 2010. Standard Methods of the International Association for Cereal Hydrocolloids, 48, 72-83.https://doi.org/10.1016/j.foodhyd.2015.02.012 Chemistry, 110/1, 105/2, 136, 104/1, 106/1, 107/1 AKIN–IDOWU, P., OYERONKE, A.E., ODUNOLA, M., YEMISI, O., IUGA, M., &MIRONEASA, S. (2019). Grape seeds effect on refined wheat flour &OLAGUNJU, Y.O. (2017). Nutritional evaluation of five species of grain dough rheology: optimal amount and particle size. Ukrainian Food Journal, 799. amaranth –An underutilized Crop. International Journal of Science, 6(1), 18-27. 10.24263/2304-974X-2019-8-4-11 DOI: 10.18483/ijSci.1131 IUGA, M., MIRONEASA, C., & MIRONEASA, S. (2019). Oscillatory rheology ALVAREZ-JUBETE, L., ARENDT, E.K., & GALLAGHER, E. (2009). and creep-recovery behaviour of grape seed-wheat flour dough: effect of grape Nutritive value and chemical composition of pseudocereals as gluten-free seed particle size, variety and addition level. Bulletin of University of ingredients. International Journal of Food Sciences and Nutrition, 60, 240-257. Agricultural Sciences and Veterinary Medicine Cluj-Napoca. Food Science and https://doi.org/10.1080/09637480902950597 Technology, 76(1), 40-51. http://dx.doi.org/10.15835/buasvmcn-fst:2018.0020 AMJID, M. R., SHEHZAD, A., HUSSAIN, S., SHABBIR, M. A., KHAN, M. R., KALINOVA, J., & DADAKOVA E. (2009). Rutin and total quercetin content in & SHOAIB, M. (2013). A comprehensive review on wheat flour dough rheology. amaranth (Amaranthus spp.). Plant Foods Human Nutrition, 64, 68–74. doi: Pakistan Journal of Food Sciences, 23(2), 105-123. 10.1007/s11130-008-0104-x. ANGIOLONI, A., &COLLAR, C. (2009). Gel, dough and fibre enriched fresh KAUR, S., SINGH, N., &RANA, J. C. (2010). Amaranthushypochondriacus and breads: Relationships between quality features and staling kinetics. Journal of Amaranthuscaudatus germplasm: Characteristics of plants, grain and flours. Food Food Engineering, 91(4), 526-532. Chemistry, 123(4), 1227-1234. https://doi.org/10.1016/j.foodchem.2010.05.091 https://doi.org/10.1016/j.jfoodeng.2008.09.033
372
J Microbiol Biotech Food Sci / Coțovanu et al. 2020 : 10 (3) 366-373
KRAUJALIS, P., & VENSKUTONIS, P.R. (2013). Optimisation of supercritical Food Science and Technology, 51(12), 4108–4113. carbon dioxide extraction of amaranth seeds by response surface methodology https://doi.org/10.1007/s13197-013-0939-5 and characterization of extracts isolated from different plant cultivars. The SANZ-PENELLA, J. M., WRONKOWSKA, M., SORAL-SMIETANA, M., & Journal of Supercritical Fluids, 73, 80–86. HAROS, M. (2013). Effect of whole amaranth flour on bread properties and https://doi.org/10.1016/j.supflu.2012.11.009 nutritive value. LWT-Food Science and Technology, 50(2), 679-685. LIU, T., HOU, G.G., LEE, B., MARQUART, L., & DUBAT, A. (2016). Effects https://doi.org/10.1016/j.lwt.2012.07.031 of particle size on the quality attributes of reconstituted whole wheat flour and SCHOENLECHNER, R., SIEBENHANDL, S., & BERGHOFER, E. (2008). tortillas made from it. Journal Cereal Science, 71, 145–152. Pseudocereals: Gluten-free cereal products and beverages 1st ed. 7, 149- https://doi.org/10.1016/j.jcs.2016.08.013 190.Academic Press. Lorenz, K. (1981). Amaranthushypochondriacus – Characteristics of the starch SINGH, N., GUJRAL, H.S., KATYAL, M., & SHARMA, B. (2019). and baking potential of the flour. Starch-Stärke, 33(5), 149–153. Relationship of Mixolab characteristics with protein, pasting, dynamic and https://doi.org/10.1002/star.19810330502 empirical rheological characteristics of flours from Indian wheat varieties with MARCIN, K., JAROSŁAW, W., MONIKA, P., & AGNIESZKA, W. (2016). diverse grain hardness. Journal of Food Science and Technology, 56, 2679–2686. Application of the response surface methodology in optimizing oat fiber particle https://doi.org/10.1007/s13197-019-03756-z size and flour replacement in wheat bread rolls. CyTA-Journal of Food, 14(1), STRUYF, N., VERSPREET, J., & COURTIN, C.M. (2016). The effect of 18–26. https://doi.org/10.1080/19476337.2015.1036309 amylolytic activity and substrate availability on sugar release in non-yeasted MARTÍNEZ M.M., DÍAZ Á., & GÓMEZ M. (2014). Effect of different dough. Journal of Cereal Science, 69, 111–118. microstructural features of soluble and insoluble fibres on gluten–free dough https://doi.org/10.1016/j.jcs.2016.02.016 rheology and bread–making.Journal of Food Engineering, 142, 49–56. SUANDARRAM, A., PANDURANGAPPA, T., & MURTHLY, K. (2014). - https://doi.org/10.1016/j.jfoodeng.2014.06.020 amylase production and applications: A review. Journal of Applied & MIRONEASA, S., & CODINĂ, G.G. (2017). The mixolab rheological properties Environmental Microbiology, 2, 166–175. DOI:10.12691/jaem-2-4-10 and dough microstructure of defatted mustard seed–wheat composite flours. SUDHA, M.L., &LEELAVATHI, K. (2012). Effect of blends of dehydrated Journal of Food Processing and Preservation, 41(5), e13130. green pea flour and amaranth seed flour on the rheological, microstructure and https://ifst.onlinelibrary.wiley.com/doi/abs/10.1111/jfpp.13130 pasta making quality. Journal of Food Science and Technology-Mysore, 49,713– MIRONEASA, S., ZAHARIA, D., CODINĂ, G.G., ROPCIUC, S., & IUGA, M. 20. DOI:10.1007/s13197-010-0213-z (2018). Effects of grape peels addition on mixing, pasting and fermentation TANG, M. C., COPELAND, L. 2007. Analysis of complexes between lipids and characteristics of dough from 480 wheat flour type. Bulletin of University of wheat starch. Carbohydrate Polymers, 67(1), 80-85. Agricultural Sciences and Veterinary Medicine Cluj-Napoca. Food Science and https://doi.org/10.1016/j.carbpol.2006.04.016 Technology, 75(1), 27-35.http://dx.doi.org/10.15835/buasvmcn-fst:0021 TESTER, R. F., KARKALAS, J., & QI, X. (2004). Starch structure and MIRONEASA, S., IUGA, M., ZAHARIA, D., & MIRONEASA, C. 2019. digestibility enzyme-substrate relationship. World’s Poultry Science Journal, Rheological analysis of wheat flour dough as influenced by grape peels of 60(2), 186-195. http://dx.doi.org/10.1079/WPS20040014 different particle sizes and addition levels. Food and Bioprocess Technology, VALCÁRCEL-YAMANI, B., & LANNES, S.D.S. (2012). Applications of 12(2), 228-245. https://doi.org/10.1007/s11947-018-2202-6 quinoa (Chenopodium quinoa Willd.) and amaranth (Amaranthus spp.) and their MIRONEASA, S., IUGA, M., ZAHARIA, D., & MIRONEASA, C. (2019a). influence in the nutritional value of cereal based foods. Food and Public Health, Optimization of grape peels particle size and flour substitution in white wheat 2(6), 265-275. DOI: 10.5923/j.fph.20120206.12 flour dough. Scientific Study & Research. Chemistry & Chemical Engineering, VENSKUTONIS, P.R., & KRAUJALIS, P. (2013). Nutritional components of Biotechnology, Food Industry, 20(1), 29-42. amaranth seeds and vegetables: a review on composition, properties, and uses. MIRONEASA, S., & MIRONEASA, C. (2019). Dough bread from refined wheat Comprehensive Reviews in Food Science and Food Safety, 12(4), 381-412. DOI: flour partially replaced by grape peels: Optimizing the rheological properties. 10.1111/1541-4337.12021 Journal of Food Process Engineering, 42(4), e13207. VITALI, D., DRAGOJEVIĆ, I.V.,&ŠEBEČIĆ, B. (2008). Extractable phenolic https://doi.org/10.1111/jfpe.13207 compounds and antioxidative properties of different wheat‐based integral MOHAMMED I., AHMED A.R., & SENGE B. (2014). Effects of chickpea flour biscuits. In: D Curic, editor. Proceedings of the 2008 Joint Central European on wheat pasting properties and bread making quality. Journal of Food Science Congress. Vol. 1. p 407– 12. and Technology, 51(9), 1902–1910. https://doi.org/10.1007/s13197-012-0733-9 WANG N., HOU G.G., KWEON M., & LEE B. (2016). Effects of particle size MOREIRA, R., CHENLO, F., TORRES, M. D., & RAMA, B. (2014). Fine on the properties of whole-grain soft wheat flour and its cracker baking particle size chestnut flour doughs rheology: Influence of additives. Journal of performance. Journal Cereal Science, 69, 187–219. Food Engineering, 120, 94-99. https://doi.org/10.1016/j.jfoodeng.2013.07.025 https://doi.org/10.1016/j.jcs.2016.03.010 MOREIRA, R., CHENLO, F., TORRES, M.D., & PRIETO D.M. (2012). WEFERS, D., TYL, C.E., & BUNZEL, M. (2015). Neutral pectin side chains of Technological Assessment of Chestnut Flour Doughs Regarding to Doughs from amaranth (Amaranthushypochondriacus) contain long, partially branched Other Commercial Flours and Formulations. Food and Bioprocess Technology, arabinans and short galactans, both with terminal arabinopyranoses. Journal of 5,2301–2310. https://doi.org/10.1007/s11947-011-0524-8 Agricultural and Food Chemistry, 63(2), 707-715. OSZVALD, M., TAMÁS, C., RAKSZEGI, M., TÖMÖSKÖZI, S., BÉKÉS, F., & https://doi.org/10.1021/jf505283x TAMÁS, L. (2009). Effects of incorporated amaranth albumins on the functional WU, C.J., & HAMADA, M.S. (2011). Experiments: planning, analysis, and properties of wheat dough. Journal of the Science of Food and Agriculture, optimization (Vol. 552). John Wiley & Sons. 89(5), 882-889. https://doi.org/10.1002/jsfa.3528 Yu, S., Ma, Y., Menager, L., & Sun, D.W. (2012). Physicochemical properties of PAHWA, A., KAUR, A., & PURI, R. (2016). Influence of hydrocolloids on the starch and flour from different rice cultivars. Food and Bioprocess Technology, quality of major flat breads: A review. Journal of Food 5(2), 626-637. https://doi.org/10.1007/s11947-010-0330-8 Processing.https://doi.org/10.1155/2016/8750258 PALOMBINI, S.V., SWAMI, T.C., MARUYAMA, A., GOHARA, A.K., SOUZA, A.H, SOUZA, N.E., VISENTAINER, J.V., GOMES, S. T., & MATSUSHITA, M. (2013). Evaluation of nutritional compounds in new amaranth and quinoa cultivars. Food Science and Technology, 33(2), 339-344. https://doi.org/10.1590/S0101-20612013005000051 REPO-CARRASCO-VALENCIA, R., HELLSTRÖM, J. K., PIHLAVA, J. M., & MATTILA, P. H. (2010). Flavonoids and other phenolic compounds in Andean indigenous grains: Quinoa (Chenopodium quinoa), kañiwa (Chenopodium pallidicaule) and kiwicha (Amaranthus caudatus). Food Chemistry, 120(1), 128- 133. https://doi.org/10.1016/j.foodchem.2009.09.087 ROSELL, C.M., CORTEZ, G., & REPO-CARRASCO, R. (2009). Breadmaking use of the Andean crops quinoa (Chenopodium quinoa), kañiwa (Chenopodiumpallidicaule), kiwicha (Amaranthuscaudatus), and tarwi (Lupinusmutabilis). Cereal Chemeistry, 86(4), 386-392. https://doi.org/10.1094/CCHEM-86-4-0386 ROSELL, C. M., COLLAR, C., & HAROS, M. (2007). Assessment of hydrocolloid effects on the thermo-mechanical properties of wheat using the Mixolab. Food Hydrocolloids, 21, 452-462. https://doi.org/10.1016/j.foodhyd.2006.05.004 SAKHARE, S. D., INAMDAR, A. A., SOUMYA, C., INDRANI, D., & RAO, G. V. (2014). Effect of flour particle size on microstructural, rheological and physico-sensory characteristics of bread and south Indian parotta. Journal of
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PROXIMATE AND CHEMICAL PROPERTIES OF SOME UNDERUTILIZED NIGERIAN WILD MUSHROOMS
Mobolaji Adenike Titilawo*1,2, Anthonia Olufunke Oluduro2, Olu Odeyemi2
Address(es): Mobolaji Adenike Titilawo, PhD 1Department of Microbiology, Osun State University, Oke-Baale, Osogbo, Nigeria. 2Department of Microbiology, Obafemi Awolowo University, Ile-Ife, Nigeria.
*Corresponding author: [email protected] doi: 10.15414/jmbfs.2020.10.3.390-397
ARTICLE INFO ABSTRACT
Received 12. 12. 2019 This investigation aims at determining the nutritional value of twenty-three underutilized wild macrofungi from a biodiversity forest in Revised 15. 7. 2020 Southwest Nigeria. The mushroom species collected across the ligneous (woody) and terrestrial (soil) habitats were analysed for Accepted 10. 8. 2020 proximate (moisture, protein, fibre, lipid, ash and carbohydrate), minerals (potassium, sodium, phosphorus, magnesium, calcium, iron Published 1. 12. 2020 and zinc) and vitamins A and C content following standard analytical procedures. Interestingly, all the mushrooms had high moisture (>80.91%) and those harvested from soil debris in the terrestrial habitat contained significantly high protein content (26.80 - 48.68%). Dietary fibre was in the range of 0.20 and 42.37%; low lipid (0.12 - 9.89%) and ash (1.25 - 14.08%) were also recorded. Furthermore, Regular article all the samples contained high carbohydrate except Macrolepiota procera (2.01%). Minerals varied across the habitats and ranged as follows: potassium (268.13 - 8972.00 mg. 100 g-1), sodium (89.36 - 425.92 mg. 100 g-1), phosphorus (0.32 - 375.51 mg. 100 g-1), magnesium (9.39 - 19.32 mg. 100 g-1) and calcium (7.98 - 37.82 mg. 100 g-1). Low iron (0.55 - 1.32 mg. 100 g-1) and zinc (2.21 - 4.98 mg. 100 g-1) were obtained. While vitamin A ranged between 0.41 and 1.41 mg. 100 g-1, vitamin C was from 4.68 to 6.93 mg. 100 g-1. Conclusively, the mushrooms investigated are a good source of nutrients and thus, can be exploited as foods or food supplements.
Keywords: wild mushrooms, habitats, nutrient composition, minerals, vitamins, Nigeria
INTRODUCTION previously reported wild Nigerian mushroom (Adeniyi et al., 2018a). In the current study, the proximate, minerals and vitamins A and C content of twenty- Mushrooms are important constituents of forest produce and very significant in three underutilized wild macrofungi from a biodiversity forest in Southwest the human diet (Gbolagade et al., 2006; Jonathan et al., 2013). They are spore- Nigeria were determined. bearing fruitbodies of fungi growing on soil or its food substrates, with some existing in natural habitats in a mycorrhizal relationship with trees (Onuoha and MATERIALS AND METHODS Obi-Adumanya, 2010), and are abundant during the rainy season in the tropics (Adeniyi et al., 2018a). Mushrooms were formerly grouped under the Kingdom Sample collection and presumptive identification Plantae that does not photosynthesize, now, classified under divisions Basidiomycota and Agaricomycota of the Fungi Kingdom (Thatoi and Exactly 10 specimens of 23 wild mushroom species were collected from the Sindevsachan, 2014). biodiversity forest of Environmental Pollution Science and Technology During the growing season of mushrooms in the wild, women, especially in rural (ENPOST) farm, Ido-Ijesa, Ilesa, Southwest Nigeria located at latitude 4°42ˈ30ˈE communities, practice traditional hunting of edible varieties (Jonathan, 2002; to 4°42ˈ45ˈˈE and longitude 7°36ˈ55ˈˈN to 7°37ˈ10ˈˈN. After harvest, Ayodele et al., 2011) for diverse purposes including nutritional and medicinal mushrooms were presumptively identified using standard keys and comparison of (Odebode, 2005; Okhuoya et al., 2010), augmenting family income the surface morphologies with existing literature (Nwordu et al., 2013; Odeyemi (Osemwegie and Okhuoya, 2009; Okhuoya et al., 2010; Ayodele et al., 2011), and Adeniyi, 2015). Following probable identification, 3 to 9 different amongst others. Generally, mushrooms are rich in proteins, vitamins and carpophores of each species were cleaned, dried and ground for further analysis. minerals, but low in cholesterol levels (Belewu and Belewu, 2005; Afiukwa et al., 2015). Factors including the growing site, substrate, and mushroom types, are Edibility test crucial to their nutrient composition (Nasiri et al., 2012; Okwulehie et al., 2014). They are considered an alternative rich source of meat, fish, vegetables, Mushrooms with characteristic woody or tough texture and small-sized were and fruits and ideal supplements for many low vitamin food materials in human classified as inedible. The soft fleshy ones were screened for the presence of diets (Afiukwa et al., 2013). toxin following the methods previously described (Feeney et al., 2014; Ohnuki Globally, the nourishment potential of the over 14,000 wild mushroom taxon’s et al., 2016). A droplet of fresh mushroom juice was placed on a piece of old has long been outshined by the well-known cultivated edible species such as paper print, allowed to dry and HCl was dropped on it. A blue spot indicated a Lentinus edodes, Pleurotus species, and Agaricus bisporus (Wasser, 2002; positive reaction for the presence of a toxin. Shelley and Geoffrey, 2004; Nakalembe et al., 2015). However, in some developing countries of the world including India, Uganda and Kenya, the dietary Proximate analysis of mushroom samples contents of some wild species are reported with much emphasis on the varieties known to be safe for consumption (Johnsy et al., 2011; Musieba et al., 2013; Proximate analysis including moisture, protein, dietary fibre, lipid, ash and Kumari and Atri, 2014; Nakalembe et al., 2009; 2015). carbohydrate of the mushrooms were performed accordingly (AOAC, 2016). In spite of the proliferation of wild mushrooms in the nooks and crannies of Nigeria, there is still a paucity of information on their nutritional values. Few Mineral determination of mushroom samples edible varieties in the genera Auricularia, Lentinus, Pleurotus, Schizophyllum, Volvoriella, Psathyrella, Russula and the highly-priced Termitomyces are Atomic absorption spectrophotometric (AAS) (Buck Scientific, 210VGP, UK) commonly reported (Egwim et al., 2011; Afiukwa et al., 2013; Ayodele and was employed in the determination of Mg, Ca, Fe and Zn using oxidizing air- Okhuoya, 2009; Ijioma et al., 2015). This represents only about 5 % of the acetylene flame at 285.2 nm, 422.6 nm, 248.3 nm, and 213.3 nm respectively,
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following the manufacturer’s instruction. Determination of sodium (Na) and others exist in mycorrhizal relationship with trees (Onuoha and Obi- potassium (K) were done using flame photometry (Jenway, PFP7, UK) following Adumanya, 2010). In the current study, eight (8) of the mushrooms were from the manufacturer’s guide. The spectrophotometry method using Yellow Vanado terrestrial habitats whereas the remaining fifteen (15) were of the ligneous origin. molybdate was used in the determination of phosphorus in the mushroom This corroborates Adeniyi et al. (2018a) who detected higher mushroom species samples (AOAC, 2016). from the ligneous habitat than the terrestrial. This could probably be attributable to the abundance of lignocellulose substrates, which support the growth of Determination of vitamins A and C mushrooms on the farm. All the terrestrial mushrooms grew directly on soil debris except Pleurotus tuber-regium, whereas the ligneous mushrooms grew on The chemicals and reagents used included HPLC-grade. Analytical reagent-grade woody substrates including decaying Cola nitida, Mangifera indica, Bambusa acetonitrile and methanol (Tedia, USA) and vitamins A and C standards (Sigma, vulgaris leaves, Cordyline australis, and Mangifera indica, decaying Elaeis Dorset) were used for the analysis. guineensis and unidentified burnt tree associated with termite nest (Tab 1).
Vitamin A Edibility of the mushroom samples
One gram (1 g) of pyrogallic acid, 70 ml ethanol and 30 ml (50 %) KOH were All the mushrooms from the terrestrial habitat were fleshy, as against those from added to 10 g mushroom powder, stirred, and refluxed for 40 min using a water the ligneous habitat that were either fleshy, woody or tough. Presumptively, nine bath (50 ± 2 ℃). Extracts were obtained three times using 50, 30 and 20 ml ether. of the mushrooms (Ganoderma applanatum, Hydnellum peckii, Hydnum sp., Double-distilled water was used to neutralize the extract, anhydrous sodium Inonotus radiatus, Marasmiellus candidus, Oxyporus populinus, Stereum sulphate added to dehydrate it and further concentrated to 5 ml using a water bath hirsutum, Trametes ochracea, and Trametes pubescens) were inedible, twelve (50 ± 2 ℃). Methanol was added to the concentrate to 10 ml mark, filtered using (Auricularia auricular-judae, Auricularia polytricha, Cantharellus cibarius, a 0.45 휇m membrane, and finally subjected to HPLC analysis. Lentinus subnudus, Macrolepiota procera, Pleurotus tuber-regium, Reversed-phase (RP) HPLC analysis was performed with the Agilent 1100 series Termitomyces striatus, Termitomyces bulborhizus, Termitomyces robustus, HPLC system (Agilent; USA), including a diode array detector. The column was Termitomyces letestui, Tricholoma inocybeoides and Tricholoma ustale) edible, made of stainless steel Agilent Eclipse XDB-C18 column (5 휇m, 4.6 × 150 mm) one (Amanita cokeri) poisonous suspect and one (Polyporus sp.) with unknown and methanol and UV detection was recorded at 325 nm for vitamin A. edibility status (Tab 1). Separation was based on isocratic elution and the solvent flow rate was maintained at 1 ml. min-1. Twenty microlitre of mushroom extract was directly injected into the HPLC column. Identification was done by comparing their retention times with those of known standards. All procedures were carried out under subdued light conditions. Standard solutions of vitamin A were prepared from stock stored in the dark at –20 ℃ by serial dilution to concentrations of 0.1, 1, 2, 5, and 10 mg per litre of vitamin A. Twenty microlitre of the standard solution was injected, and peak areas were determined to generate standard curves.
Vitamin C
Ten grams (10 g) of mushroom powder was mixed with extracting solution prepared by dissolving 15 g of metaphosphoric acid with 40 ml acetic acid and made up to 500 ml with distilled water. The mixture was filtered through Whatman No. 1 filter paper and samples were extracted in triplicate. Ascorbic a b acid standard solution was prepared by weighing 100 mg ascorbic acid, dissolved and made up to 100 ml with metaphosphoric acid – acetic solution. The calibration line was converted to a line arrange based on four measured concentration levels. Quantification of ascorbic acid content was performed on an Agilent HPLC system. Chromatographic separation was achieved on an RP- HPLC column through the isocratic delivery of a mobile phase (A/B 33/67; A: 0.1M potassium acetate, pH = 4.9, B: acetonitrile: water [50:50]) at a flow rate of 1 ml. min-1. UV absorbance was recorded at 254 nm at room temperature.
Data analysis
All experiments were carried out in triplicates. Data analyses were performed using SPSS IBM version 23 software. All data were expressed as mean ± standard deviation. The difference in means was evaluated using ANOVA and c d Duncan multiple tests employed in case of variance heterogeneity. Values with a probability level of less than 0.05 were considered significant. Nutritional distance and similarity among mushrooms were determined and dendrogram obtained using a complete linkage approach.
RESULTS AND DISCUSSION
Presumptive identification of mushroom samples
Mushrooms play significant roles in biogeochemical recycling of elements in the environment, human nutrition and dietetics, and medicines (Odeyemi et al., 2014). In this study, a total of 23 different mushroom samples were collected between April and October, 2018 at ENPOST farm, Ido-Ijesa, Ilesa, Southwest Nigeria, and classified into 17 genera, namely: Amanita, Auricularia, e f Cantharellus, Ganoderma, Hydnellum, Hydnum, Inonotus, Lentinus, Macrolepiota, Marasmiellus, Oxyporus, Pleurotus, Polyporus, Stereum, Termitomyces, Trametes and Tricholoma. Representative pictures of the mushrooms are shown in figure 1. All the genera had one species each except Auricularia (2), Termitomyces (4), Trametes (2) and Tricholoma (2). These mushrooms had been previously reported (Zoberi, 1973; Alofe, 1985; Gbolagade et al., 2006; Oyetayo, 2009; Okhuoya et al., 2010; Adedayo, 2011; Djelloul and Samraoul, 2011; Johnsy et al., 2011; Nwordu et al., 2013; Adeniyi et al., 2018a;b). In nature, mushrooms grow on lignocellulose containing substrates (Jonathan et al., 2013; Adeniyi et al., 2018a). While some grow on soil or wood substrates,
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g h i j Figure 1 Representative pictures of the mushroom samples Legend: a - Auricularia polytricha, b - Amanita cokeri, c - Ganoderma applanatum, d - Inonotus radiatus, e - Macrolepiota procera, f - Marasmiellus candidus, g - Oxyporus populinus, h - Polyporus sp., i - Termitomyces bulborhizus, j - Trametes ochracea
Table 1 Characteristics of mushrooms used in the study S/N Mushroom Habitat Substrate Texture Edibility Date of collection 1 Amanita cokeri Terrestrial Soil debris Fleshy Poisonous / suspect 05/04/2019 2 Auricularia auricular-judae Ligneous Dead Cola nitida stem log Fleshy Edible 29/05/2019 3 Auricularia polytricha Ligneous Dead Mangifera indica stem log Fleshy Edible 21/06/2019 4 Cantharellus cibarius Ligneous Decaying Elaeis guineensis Fleshy Edible 17/04/2019 5 Ganoderma applanatum Ligneous Dead Cola nitida stem log Woody Inedible 19/05/2019 6 Hydnellum peckii Ligneous Bambusa vulgaris leaves Tough Inedible 14/05/2019 7 Hydnum sp. Ligneous Cordyline australis tree Tough Inedible 25/04/2019 8 Inonotus radiatus Ligneous Dead Mangifera indica stem log Woody Inedible 16/06/2019 9 Lentinus subnudus Ligneous Dead Mangifera indica stem log Fleshy Edible 23/04/2019 10 Macrolepiota procera Terrestrial Soil debris Fleshy Edible 10/05/2019 11 Marasmiellus candidus Ligneous Decaying Elaeis guineensis Fleshy Inedible 13/05/2019 12 Oxyporus populinus Ligneous Dead Cola nitida stem log Woody Inedible 19/05/2019 13 Pleurotus tuber-regium Terrestrial Sclerotium on soil debris Fleshy Edible 02/05/2019 14 Polyporus sp. Ligneous Dead Cola nitida stem log Fleshy Unknown 08/06/2019 15 Stereum hirsutum Ligneous Dead Mangifera indica stem log Tough Inedible 20/05/2019 Unidentified burnt tree associated 16 Termitomyces striatus Ligneous Fleshy Edible 12/06/2019 with termite nest 17 Termitomyces bulborhizus Terrestrial Termite mound Fleshy Edible 09/10/2019 18 Termitomyces robustus Terrestrial Termite mound Fleshy Edible 14/10/2019 19 Termitomyces letestui Terrestrial Termite mound Fleshy Edible 12/10/2019 20 Trametes ochracea Ligneous Dead Cola nitida stump Woody Inedible 25/08/2019 21 Trametes pubescens Ligneous Dead Mangifera indica stem log Woody Inedible 25/08/2019 22 Tricholoma inocybeoides Terrestrial Soil debris Fleshy Edible 07/05/2019 23 Tricholoma ustale Terrestrial Termite nest Fleshy Edible 25/08/2019
Proximate analysis of the mushroom samples detected in Hydnum sp. (11.48 ± 0.06 %). Protein obtained for other edible species including A. auricular-judae (19.60 ± 0.06%), A. polytricha (12.29 ± Moisture contents 0.06%), L. subnudus (18.74 ± 0.12%) and P. tuber-regium (19.84 ± 0.67%) were higher than the report of Johnsy et al. (2011) but lower than the findings of Water exists virtually in all foods, and its importance in many physiological Gbolagade et al. (2006). The difference in geographical location and substrates processes cannot be over-emphasized. Generally, all the mushrooms analysed in may be responsible for this. our study contained high moisture ranging from 80.91 to 98.44% (Tab 2) and agrees with Gbolagade et al. (2006) and Johnsy et al. (2011) whose report Dietary fibres ranged from 85.4 to 98.5 % and 87.3 to 95.17% respectively. High moisture contents in mushrooms contribute to low shelf life owing to high water activity Current trends recommend a drive towards diets that contain a greater amount of that enhances microbial growth and enzyme activity (Gbolagade et al., 2006; plant foods as implicated in preserving and/or improving personal wellbeing Johnsy et al., 2011). Moisture variation may be linked to prevailing (Rodrı´guez et al., 2006; Usha and Suguna, 2014). Dietary fibres of the environmental growth conditions (Mattila et al., 2001). mushrooms studied varied across habitats (Tab 2). With the exception of T. ochracea (0.20 ± 0.01 %) and G. applanatum (42.37 ± 0.02%), the values Proteins recorded fell within the ranges reported elsewhere (Breene, 1990; Chye et al., 2008; Egwim et al., 2011; Johnsy et al., 2011; Musieba et al., 2013; Kumari Protein contents of mushrooms vary according to the genetic make-up of species and Atri, 2014). The significantly high dietary fibre in G. applanatum indicates and disparity in physicochemical properties of growth substrate (Ragunathan the presence of high leftovers of plant cells resistant to hydrolysis by the and Swaminathan, 2003; Sanmee et al., 2003; Agrahar-Murugkar and alimentary enzymes of man (Capuano, 2017). However, the value obtained in T. Subbulakshmi, 2005). Overall, mushrooms harvested from soil debris (terrestrial ochracea is comparable with broccoli (0.15%), red radish (0.19%), white radish habitat) were higher in protein than those from the ligneous habitat (Tab 2). This (0.16%), sweet yellow pepper (0.11%), spring onions (0.21%) and tomato may be due to high organic matter in substrates constituting the habitat. Of all the (0.18%) (Januškevičius et al., 2012). Dietary fibre benefits in preventing of higher fungi investigated, M. procera was the richest in protein (48.68 ± 0.13%). heart diseases, colon cancer and diabetes (Kassegn, 2018). This disagrees with Kumari and Atri (2014) who reported 19.95 ± 1.06% for the same mushroom. The protein value (45.50 ± 0.06%) obtained for A. cokeri, a Lipids poisonous suspect mushroom was higher than the presumed edible Termitomyces and Tricholoma species suggesting it as an attractive and useful species for Lipids provide the major caloric value in foods and excess intake can biotechnological manipulation as a food source. The lowest protein content was consequentially lead to coronary heart disease and other health issues (dos
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Passos et al., 2013). Mushrooms are low in lipids dominated by unsaturated fatty acids, contributing less to the energy required in human diet and serving as a Carbohydrates means of combating obesity (Wani et al., 2010). Generally, the current investigation ranged between 0.12 and 9.89% (Tab 2). This tally with the Carbohydrates are good sources of energy desirable for breakfast and weaning observation of Maftoun et al. (2015). Difference in mushroom species may food formulae (Kassegn, 2018). In mushrooms, carbohydrate content constitutes account for the varied lipid contents observed in the study. the bulk of fruiting bodies accounting for 50 to 65% on a dry weight basis (Wani et al., 2010; Di Aniba et al., 2015). In the present investigation, carbohydrate Ash ranged from 2.01 to 67.73% with M. procera and T. pubescens having the lowest and highest values of 2.01 ± 0.04% and 67.44 ± 0.06% respectively (Tab 2). Our Ash contents indicate the presence of mineral contents in any given food sample findings on M. procera concur with Egwim et al. (2011) who indicated 8% but (Kassegn, 2018). Other than T. pubescens (0.64 ± 0.01%), T. inocybeoides (1.25 opposes Kumari and Atri (2014) who reported 60.82%. Similarly, Usha and ± 0.01%) and T. ustale (1.37 ± 0.01%), ash contents obtained for others Suguna (2014) reported lower carbohydrate content (28.5 %) for A. polytricha in correspond with the outcomes of Gbolagade et al. (2006) and Colak et al. their study. In the case of Johnsy et al. (2011), 50.20, 43.4, 47.83 and 33.23% (2009) who had 4.7 to15.5% and 2.00 to 11.38% respectively (Tab 2). Previous were documented for P. tuber-regium, P. ostreatus, Lentinus squarrosulus and A. study reported relatively high ash contents in Termitomyces microcarpus auricular-judae respectively. (15.13%) and Termitomyces clypeatus (16.87%) (Nakalembe et al., 2015).
Table 2 Proximate composition of the twenty-three mushrooms (dry weight basis) Mushroom MC P DF LD A CHO [%] [%] [%] [%] [%] [%] Amanita cokeri 95.84±0.08efg 45.50±0.06b 12.44±0.01h 3.81±0.01l 12.32±0.01c 21.50±0.62q Auricularia auricular-judae 95.55±0.30fgh 19.60±0.06jk 2.41±0.02r 0.53±0.03r 4.98±0.02k 67.40±0.04a Auricularia polytricha 94.92±0.68ij 12.29±0.06q 4.32±0.01t 0.55±0.01r 2.27±0.01r 67.44±0.06a Cantharellus cibarius 95.94±0.09ef 31.05±0.06f 4.56±0.01s 9.30±0.01b 11.53±0.03e 29.94±0.04o Ganoderma applanatum 95.12±0.28hi 21.20±0.12i 42.37±0.02a 6.60±0.01e 2.09±0.01s 17.64±0.12r Hydnellum peckii 94.31±0.01k 21.40±0.00i 18.55±0.03c 1.14±0.01o 4.57±0.01m 43.94±0.01i Hydnum sp. 91.25±0.07n 11.48±0.06r 7.34±0.01o 6.09±0.01g 14.02±0.03b 48.18±0.08g Inonotus radiatus 80.91±0.13q 16.20±0.06m 12.41±0.02i 0.93±0.01q 11.88±0.01d 44.96±0.03h Lentinus subnudus 95.31 ±0.44ghi 18.74±0.12l 29.91±0.01b 1.00±0.01p 14.08±0.02a 27.73±0.12p Macrolepiota procera 95.59±0.16efgh 48.68±0.13a 14.35±0.01e 8.59±0.02c 10.83±0.02f 2.01±0.04s Marasmiellus candidus 94.54±0.28jk 31.57±0.06e 8.38±0.01m 6.48±0.02f 7.69±0.01h 38.14±0.05l Oxyporus populinus 88.87±0.19 o 19.44±0.06k 11.77±0.01k 0.12±0.01u 4.77±0.01l 51.90±0.08e Pleurotus tuber-regium 95.41±0.28fghi 19.84±0.67j 6.45±0.01p 0.26±0.01t 2.62±0.01o 55.91±0.64d Polyporus sp. 92.45±0.06m 15.30±0.00n 13.03±0.00g 0.44±0.01s 2.34±0.01q 56.27±0.03cd Stereum hirsutum 83.09 ±0.13p 14.67±0.00o 13.25±0.01f 1.97±0.01m 2.43±0.02p 56.48±0.03c Termitomyces striatus 94.91 ± 0.14ij 26.53±0.12h 12.36±0.01j 7.44±0.01d 4.76±0.01l 42.69±0.43j Termitomyces bulborhizus 98.44±0.20a 27.53±0.06g 6.25±0.01q 4.74±0.03k 5.97±0.01j 49.00±0.01f Termitomyces robustus 97.80±0.15b 31.00±0.06f 7.36±0.01o 6.50±0.02f 4.36±0.01n 39.51±0.08k Termitomyces letestui 96.95±0.08c 26.80±0.06h 5.46±0.14r 5.21±0.14i 7.45±0.01i 48.80±0.06f Trametes ochracea 93.53 ±0.20l 16.04±0.06m 0.20±0.01u 5.43±0.01h 9.26±0.01g 57.27±0.06b Trametes pubescens 94.10 ± 0.13k 12.77±0.06p 16.61±0.01d 1.82±0.01n 0.64±0.01v 67.73±0.08a Tricholoma inocybeoides 96.10±0.13de 41.30±0.06c 9.21±0.01l 4.92±0.01j 1.25±0.01u 31.45±0.06n Tricholoma ustale 96.46±0.20cd 34.43±0.12d 7.53±0.01n 9.89±0.01a 1.37±0.01t 33.48±0.04m Mean with a different letter in each column are significantly different from each other at p < 0.05. Mean values are averages of 3 values. Legend: MC - Moisture content, P - Protein, DF - Dietary fibre, LD - Lipid, A - Ash, CHO - Carbohydrate.
Mineral and vitamin compositions of mushroom samples stones and severity of asthma symptoms (de Wardener and MacGregor, 2002; Mohan et al., 2009). Potassium Phosphorus Potassium controls the water and minerals in the blood and tissues, and significant in the transmission of electrical impulses in the heart (Kowey, 2002). Phosphorus is needed to build strong and healthy bones, sustains normal Among all the nutrients analysed, potassium predominates other minerals (Tab acid/base balance, supports growth and is involved with the storage and use of 3). This is in line with some studies conducted elsewhere (Akyüz and Kirbağ, energy (dos Passos et al., 2013). In this study, phosphorus was found to vary 2010; Ayodele and Okhuoya, 2009; Okwulehie and Ogoke, 2013; Nakalembe across habitats and differ significantly (p < 0.05). Its content (0.32 - 375.51 mg. et al., 2015). However, the range of potassium indicated in this study (268.13 - 100 g-1) (Tab 3) was lower than the report of Nakalembe et al. (2015) (10.2 - 8972.00 mg. 100 g-1) was higher than those of Gbolagade et al. (2006) (7.8 - 16.8 mg. 100 g-1) but higher in Okwulehie and Ogoke (2013) except for 60.1 mg. 100 g-1) and Okwulehie and Ogoke (2013) (112.14 - 164.54 mg. 100 g- Hydnum sp. (23.62 mg. 100 g-1) and Trametes pubescens (0.32 mg. 100 g-1) (Tab. 1). 3). The tolerable upper intake level for phosphorus is 3 µg per day for children and 3 to 4 µg per day for adults (Food and Nutrition Board, 2016) suggesting Sodium that little dose of the mushrooms is needed in daily diet.
Sodium is the main electrolyte and major cation outside the cell (Knochel, 1999; Magnesium Wardlaw and Kessel, 2002). In contrast to some previous studies (Gbolagade et al., 2006; Akyüz and Kirbağ, 2010), the sodium content of mushrooms studied Magnesium ranged from 9.39 to 19.32 mg. 100 g-1) (Tab 3) and its lower was significantly high (89.36 - 425.92 mg. 100 g-1) (Tab 3). While Gbolagade et compared to many common legumes (178 - 197 mg. 100 g-1), whole-grain cereal al. (2006) documented a range of 0.2 to 6.3 mg. 100 g-1, Akyüz and Kirbağ (121 - 434 mg. 100 g-1) (FAO, 2012), and edible mushrooms (Nakalembe et al., (2010) had 0.2 to 1.2 mg. 100 g-1. Furthermore, the values obtained exceeded the 2015). However, it is higher than Sanmee et al. (2003) and Gbolagade et al. recommendations of the World Health Organisation (WHO) daily sodium intake (2006) who observed a range of 0.5 - 1.6 mg. 100 g-1 and 0.4 - 6.7 mg. 100 g-1 (≤2000 mg) and the UK government (2400 mg per day) (WHO, 2006; Mohan et respectively. Magnesium is crucial in control of muscle contraction, blood al., 2009). High dietary sodium has been found implicated in some health pressure, insulin metabolism, and synthesis of DNA, RNA and proteins (Grober consequences including an increase in blood pressure, hypertension, direct et al., 2015). It plays an important role in nerve transmission and neuromuscular vascular and cardiac damage, obesity, stomach cancer, osteoporosis, kidney conduction and protection against excessive excitation that can lead to excitotoxicity (Kirkland et al., 2018).
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Calcium stunted growth and hypogonadism. Low levels of zinc can as well weaken the body’s immunity (Edward et al., 2013; Titilawo et al., 2018). Calcium is one of the foremost microminerals in the bone and teeth of human beings and animals and reduces the chance of having cardiac disorder or any Vitamin A heart-related challenge (Titilawo et al., 2018). The obtained values (7.98 - 37.82 mg. 100 g-1) (Tab 3) were comparable to many common kinds of cereals (12 - 51 Mushrooms are non-animal sources of vitamin, contributing a very small mg. 100 g-1), starchy food (11 - 43 mg. 100 g-1), meat and poultry products (4 - percentage in human diet and crucial in disease prevention and lengthening of life 69 mg. 100 g-1) (FAO, 2012). However, Chye et al. (2008) obtained a range (77 - span (Olaniyi, 2000). The outcome of our work revealed that all the wild 144.7 mg. 100 g-1) higher than ours. Consumption of a very high concentration of mushrooms analysed contained vitamin A (Tab 3). This is contrary to Afiukwa et calcium may adversely affect the absorption of the essential elements (Nova al. (2013) who detected vitamin A only in Agaricus bisporus among other Scotia Environment, 2005), nonetheless, its paucity could result in osteoporosis varieties and with a value higher (38.36 mg. 100 g-1) than what was observed in and osteomalacia. Hypertension has also been linked to low calcium in the body our case (0.14 - 1.41 mg. 100 g-1). Likewise, Musieba et al. (2013) investigated (Kožĺšek, 2003; Titilawo et al., 2018). vitamin A in P. citrinopileatus and reported a low value <10 µg/100 g. Our findings agree with vitamin A level reported for some animal and plant materials Iron including butter (0.59 mg. 100 g-1), cheese (0.39 mg. 100 g-1), egg (0.28 mg. 100 g-1), milk (0.04 mg. 100 g-1) and salmon (0.041 mg. 100 g-1) (Souci et al., 2000). Iron is required for the synthesis of haemoglobin in red blood corpuscles and also Vitamin A role has been established to include apt sight, reproduction, growth aids growth and metabolic processes in humans and animals (Kumar and Puri, and development, cellular differentiation and immune function (Bowman, 2001). 2012, Edward et al., 2013; Vetrimurugan et al., 2017; Titilawo et al., 2018). In this work, the iron content (0.55 - 1.30 mg. 100 g-1) (Tab 3) was within the Vitamin C published data for cabbage (0.6 mg. 100 g-1) and meat (1.6 mg. 100 g-1) (Eyabi, 2001). It was however higher than those observed in previous studies The range of vitamin C obtained in this work was from 5.22 to 6.93 mg. 100 g-1. (Gbolagade et al., 2006; Ayodele and Okhuoya, 2009), and lower when While the highest vitamin C content was found in Pleurotus tuber-regium, the compared with the findings of Colak et al. (2009), Patil et al. (2010) and lowest was detected in T. pubescens (Tab. 3). The values indicated that the Kumari and Atri (2014). Largely, a low quantity of iron was observed in the mushrooms are considered good sources of ascorbic acid. Earlier studies by mushrooms and fell below the recommended nutritional limit (18 mg) for iron, Sapers et al. (1999), Mattila et al. (2001) and Mattila et al. (2000) reported (Food and Nutrition Board, 2016). Thus, the mushrooms are not good sources vitamin C content of up to 7 mg. 100 g-1. Also, Muthangya et al. (2014) of dietary iron. observed 5.07 to 5.29 mg. 100 g-1 in edible sections of mushrooms whereas Musieba et al. (2013) recorded a low value of <1 µg. 100 g-1 in Pleurotus Zinc citrinopileatus. Ascorbic acids are major antioxidants with known health benefits (Rao and Agarwal, 2000). It empowers the body against viral and bacterial Zinc is a major micronutrient for living organisms. It is present almost in all attack and also vital in wound healing (Hemilä, 2017). It is vital for the foods and water as a salt of organic complexes (Swaminathan et al., 2011; production of collagen, a key structural constituent of tendons, bones, teeth, Salano, 2013). Generally, the mushrooms in the current study are low in zinc and blood vessels and muscles. Vitamin C enhances iron absorption and regenerates fell below the 15 mg recommended dietary intake (Food and Nutrition Board, other antioxidants including vitamin E (Brody, 1994). On the contrary, 2016). The range obtained was between 2.21 and 4.98 mg. 100 g-1 (Tab 3). This deficiency in vitamin C causes bruising, haemorrhage, dry skin and depression is lower than the results of previous studies (Gbolagade et al., 2006; Nakalembe (Olson, 1999). The vitamin is reported to directly interact with radicals in et al., 2015) but higher than the report of Okwulehie and Ogoke (2013). When plasma, averting impairment to red cell membranes (Jayakumar et al., 2009). zinc is lacking in the diet, it may lead to a reduction in fertility, forfeiture of taste,
Table 3 Mineral and vitamin composition of the selected mushroom samples (dry weight basis) (mg. 100g-1) Mushroom Sodium Potassium Magnesium Calcium Iron Zinc Phosphorus Vitamin A Vitamin C Amanita cokeri 280.37±0.01f 8972.00±0.05a 12.68±0.01n 28.04±0.01e 1.05±0.01f 2.78±0.01l 127.56±0.01l 1.14±0.01ef 5.48±0.02cd Auricularia auricular-judae 131.57±0.02s 2473.68±0.01k 15.36±0.01l 8.76±0.01t 1.22±0.01bc 2.33±0.01n 159.55±0.02i 1.34±0.01b 6.25±0.01b Auricularia polytricha 231.56±0.01h 1260.72±0.01o 18.56±0.01d 17.14±0.01j 1.01±0.01g 2.32±0.01n 93.25±0.01m 1.23±0.01c 5.34±0.01cd Cantharellus cibarius 425.15±0.25b 5954.62±0.01c 15.36±0.01l 37.82±0.01a 1.24±0.01b 3.87±0.02b 208.35±0.01g 1.12±0.01fg 6.43±0.02b Ganoderma applanatum 152.43±0.02q 670.73±0.00u 17.69±0.01e 10.17±0.01l 1.30±0.01a 3.25±0.01j 75.74±0.02r 1.22±0.01cd 5.44±0.01cd Hydnellum peckii 196.25±0.02k 1570.10±0.01m 10.36±0.01p 18.67±0.02h 1.30±0.01a 3.49±0.01f 90.60±0.01n 1.23±0.01c 6.32±0.01b Hydnum sp. 174.43±0.01m 406.97±0.01v 19.32±0.01a 29.08±0.01d 1.10±0.02e 2.21±0.01o 23.62±0.01v 1.41±0.01a 5.38±0.01cd Inonotus radiatus 247.26±0.01g 1236.28±0.02p 17.67±0.01e 36.65±0.02b 1.32±0.01a 2.54±0.01m 80.99±0.01p 1.11±0.01fg 6.22±0.01b Lentinus subnudus 113.11±0.01u 1724.78±0.01l 16.32±0.02h 9.32±0.11q 0.64±0.01k 3.13±0.01k 154.83±0.01j 1.36±0.01b 5.56±0.04c Macrolepiota procera 166.35±0.01o 3632.14±0.04h 15.84±0.01j 9.24±0.02r 0.74±0.02i 3.25±0.13j 70.07±0.01u 1.04±0.01i 6.32±0.01b Marasmiellus candidus 329.97±0.05c 4241.96±0.01f 18.92±0.01b 31.43±0.01c 1.24±0.01b 3.44±0.01g 375.51±0.37a 1.32±0.01b 6.39±0.02b Oxyporus populinus 199.44±0.01i 1453.00±0.01n 17.67±0.01e 9.51±0.01o 0.82±0.01h 3.63±0.01d 193.94±0.00h 1.05±0.07hi 5.39±0.02cd Pleurotus tuber-regium 115.95±0.01t 927.55±0.01t 16.29±0.01h 19.31±0.01g 0.68±0.01j 3.44±0.01g 80.29±0.01q 1.09±0.01gh 6.93±0.54a Polyporus sp. 158.03±0.01p 1106.22±0.02r 10.32±0.01q 17.56±0.01i 1.30±0.01a 3.55±0.01e 74.12±0.01s 1.02±0.01i 5.29±0.02cd Stereum hirsutum 93.01±0.01v 953.49±0.00s 9.39±0.02r 15.52±0.02k 1.13±0.02d 3.34±0.01i 73.63±0.01t 1.18±0.04de 6.22±0.01b Termitomyces striatus 195.34±0.01l 5860.46±0.01d 16.49±0.02g 9.32±0.02q 0.69±0.01j 3.78±0.01c 371.89±0.01b 1.12±0.01fg 6.29±0.02b Termitomyces bulborhizus 167.07±0.01n 5250.60±0.01e 15.70±0.01k 7.98±0.02v 0.55±0.01l 4.98±0.01a 222.55±0.01e 1.23±0.01c 6.36±0.04b Termitomyces robustus 198.01±0.01j 7673.26±0.01b 15.29±0.02m 8.24±0.02u 1.09±0.01e 3.37±0.01i 217.15±0.01f 1.23±0.01c 5.49±0.01c Termitomyces letestui 310.43±0.01d 3668.87±0.01g 18.78±0.01c 9.42±0.01f 1.19±0.01c 3.40±0.01h 271.50±0.01c 1.14±0.01ef 6.43±0.01b Trametes ochracea 138.13±0.01r 1160.24±0.02q 11.66±0.01o 27.63±0.01f 1.02±0.01g 3.53±0.01e 86.64±0.01o 0.14±0.01j 5.22±0.01cd Trametes pubescens 89.36±0.01w 268.13±0.01w 16.26±0.02i 9.94±0.01m 1.01±0.01g 2.55±0.01m 0.32±0.01w 1.33±0.01b 4.68±0.01e Tricholoma inocybeoides 425.92±0.01a 2608.70±0.00j 15.32±0.01m 8.84±0.05s 1.21±0.01bc 3.36±0.01i 146.71±0.01k 1.19±0.01cd 6.29±0.01b Tricholoma ustale 292.96±0.01e 3281.11±0.21i 16.67±0.02f 9.77±0.01n 1.14±0.00d 3.39±0.01h 223.34±0.01d 1.12±0.01fg 5.48±0.01cd Mean with a different letter in each column are significantly different from each other at p < 0.05. Mean values are averages of 3 values.
Nutritive hierarchical cluster analysis of the mushrooms (figure 2). For instance, the fleshy, edible P. tuber-regium and T. inocybeoides in the terrestrial habitat closely associated with woody/tough, inedible G. Mushrooms fruitbodies are characterized by assimilated mineral constituents applanatum and S. hirsutum in the ligneous habitat respectively. Likewise, the whose level depends on, amongst other species, age of mushroom, the diameter fleshy and edible T. striatus and C. cibarius in the ligneous habitat closely related of pileus and substratum (Dermirdaş, 2001; Falandysy et al., 2001; Mattila et to fleshy, edible T. bulborhizus (terrestrial habitat); and the fleshy, inedible M. al., 2001). The nutrition hierarchy cluster shows that there is dietary relatedness candidus in the ligneous habitat found clustered with fleshy, edible M. procera, among the mushroom species irrespective of their habitat, texture or edibility T. ustale and T. letestui in the terrestrial habitat (figure 2). Also, the nature of the
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nutrient-rich organic substrate on which M. candidus, C. cibarius, and T. striatus Adeniyi, M.A., Odeyemi, Y. & Odeyemi, O. (2018a). Ecology, diversity and grew may be responsible for dietary relatedness with mushrooms from terrestrial seasonal distribution of wild mushrooms in a Nigerian tropical forest reserve. habitat. Likewise, the presence of sclerotium in P. tuber-regium and decaying Biodiversitas 19(1), 285-295. https://doi.org/10.13057/biodiv/d190139 debris where Tricholoma inocybeoides were harvested may be the reason their Adeniyi, M.A., Titilawo, Y., Oluduro, A.O., Odeyemi, O., Nakin, M. & Okoh, nutrient content is comparable with mushrooms from the ligneous habitat. A.I. (2018b). Molecular identification of some wild Nigerian mushrooms using internal transcribed spacer: polymerase chain reaction. AMB Express 8, 148. https://doi.org/10.1186/s13568-018-0661-9 Afiukwa, C. A., Ebem, E. C. & Igwe, D. O. (2015). Characterization of proximate and amino acid composition of edible wild mushroom species in Abakaliki, Nigeria. America Association for Science and Technology 1(2), 20-25. Afiukwa, C.A., Ugwu, O.P.C., Okoli, S.O., Idenyi, J.N. & Ossai, E.C. (2013). Contents of some vitamins in five edible mushroom varieties consumed in Abakaliki metropolis, Nigeria. Research Journal of Pharmaceutical, Biological and Chemical Sciences 4(2), 805-812. Agrahar-Murugkar, D. & Subbulakshmi, G. (2005). Nutritional value of edible wild mushrooms collected from the Khasi hills of Meghalaya. Food Chemistry 89, 599-603. https://doi.org/10.1016/j.foodchem.2004.03.042 Akyüz, M. & Kirbağ, S. (2010). Nutritive value of wild edible and cultured mushrooms. Turkish Journal of Biology 34, 97-102. https://doi.org/10.3906/biy- 0805-17 Alofe, F.V. (1985). The general characteristics and cultivation of some Nigerian mushrooms. Ph.D. Thesis, Obafemi Awolowo University, Ile-Ife, Nigeria. AOAC (2016). Official methods of analysis of the association of official analytical chemists. 20th ed., Washington, DC. Ayodele, S. M., Akpaja, E. O. & Adamu, Y. (2011). Some edible medicinal mushrooms of Igala Land in Nigeria, their socio-cultural and ethnomycological uses. International Journal of Science and Nature 2(3), 473-476. Ayodele, S.M. & Okhuoya, J.A. (2009). Nutritional and phytochemical evaluation of cultivated Psathyrella atroumbonata Pegler, a Nigerian edible mushroom. South African Journal of Science 105, 158-160. https://doi.org/10.4102/sajs.v105i3/4.72 Belewu, M. A. & Belewu, K. Y. (2005). Cultivation of mushroom (Volvariella volvacea) on banana leaves. African Journal of Biotechnology 4(12), 1401-1403. Beluhan, S. & Ranogajec, A. (2011). Chemical composition and nonvolatile components of Croatian wild edible mushrooms. Food Chemistry 124, 1076- Figure 2 Dendrogram of the nutritive value of the selected mushroom samples 1082. https://doi.org/10.1016/j.foodchem.2010.07.081 Legend: 1 - Macrolepiota procera, 2 - Amanita cokeri, 3 - Tricholoma inocybeoides, 4 - Tricholoma ustale, 5 - Termitomyces robustus, 6 - Termitomyces bulborhizus, 7 - Bowman, B.A. & Russell, R.B. (2001). Present knowledge in nutrition. Termitomyces letestui, 8 - Pleurotus tuber-regium, 9 - Marasmiellus candidus, 10 - Washington, DC: International Life Science Institute, Press. Cantharellus cibarius, 11 - Termitomyces striatus, 12 - Hydnellum peckii, 13 - Hydnum sp., Breene, W.M. (1990). Nutritional and medicinal value of specialty mushrooms. 14 - Ganoderma applanatum, 15 - Auricularia auricular-judae, 16 - Oxyporus populinus, 17 Journal of Food Product 53(10), 883-894. - Trametes ochracea, 18 - Polyporus sp., 19 - Inonotus radiatus; 20 - Stereum hirsutum, 21 - Brody, T. (1994). Nutritional biochemistry. San Diego, CA: Academic Press. Trametes pubescens, 22 - Lentinus subnudus, 23 - Auricularia polytricha Capuano, E. (2017). The behavior of dietary fiber in the gastrointestinal tract determines its physiological effect. Critical Reviews in Food Science and CONCLUSION Nutrition 57(16), 3543-3564. https://doi.org/10.1080/10408398.2016.1180501 Chye, F. Y., Wong, J. Y. & Lee, J. S. (2008). Nutritional quality and antioxidant The nutritional composition of twenty-three underutilized Nigerian wild activity of selected edible wild mushrooms. Revista de Agaroquimica y mushrooms was studied. While eight of the mushrooms were from the terrestrial Tecnologia de Alimentos 14(4), 375-384. habitat, fifteen were of the ligneous source. Interestingly, all the mushroom https://doi.org/10.1177%2F1082013208097445 species are rich in moisture, protein, dietary fibre, ash, carbohydrate and minerals Colak, A., Kolcuoğlu, Y., Sesli, E. & Dalman, Ö. (2009). Biochemical including sodium, potassium, calcium, magnesium, zinc, and phosphorus, and composition of some Turkish fungi. Asian Journal of Chemistry 19, 2193-2199. vitamins A and C, but low in lipid and iron, an indication that they are highly de Wardener, H.E. & MacGregor, G.A. (2002). Harmful effects of dietary salt in nutritious; although other sources of sodium must be checked when consuming addition to hypertension. Journal of Human Hypertension 16, 213-223. these mushrooms. Similarly, the nutritive hierarchical cluster analysis shows that https://doi.org/10.1038/sj.jhh.1001374 the dietary composition of all the mushrooms is related irrespective of their Dermirbaş, A. (2001). Concentrations of 21 metals in 1 species of mushrooms habitat, texture or edibility. Thus, they are potential foods and food supplements growing in the East Black Sea region. Food Chemistry 75, 453-457. and stand to meet the diverse human dietary needs if objectively exploited. Di Anibal, C., Farenzena, S., Rodrı´guez, M.S. & Albertengo, L. (2015). Further study is however encouraged to ascertain the safety of these mushrooms Chemical composition and nutritional value of Argentine commercial edible for consumption. mushrooms Journal of Consumer Protection and Food Safety 10, 155-164. https://doi.org/10.1007/s00003-015-0937-9 Conflicts of interest: The authors declare no conflict of interest. Djelloul, R. & Samraoui, B. (2011). Distribution and ecology of the superior mushrooms of the Aulnaie of Ain Khiar (El Kala National Park, Northeastern Author contributions: M.A.T and O.O. designed the experiment, M.A.T. Algeria). African Journal of Environmental Science and Technology 5(6), 448- performed the experiment and drafted the manuscript, M.A.T. and A.O.O. 456. analysed the data, A.O.O. and M.A.T. supervised the study. All authors read and dos Passos, M.E.A, Moreira, C.F.F., Pacheco, M.T.B., Takase, I., Lopes, M.L.M. approved the final manuscript. & Valente-Mesquita, V.L. (2013). Proximate and mineral composition of industrialized biscuits. Food Science and Technology 33(2), 323-331. Acknowledgement: This research did not receive any specific grant from http://dx.doi.org/10.1590/S0101-20612013005000046 funding agencies in the public, commercial or not-for-profit sectors. However, Edward, J.B., Idowu, E.O., Oso, J.A. & Ibidapo, O.R. (2013). Determination of the authors appreciate the management of ENPOST farm, Ido-Ijesa, Ilesa, heavy metal concentration in fish samples, sediment and water from Odo-Ayo Nigeria for permission to collect mushroom samples. Special thanks goes to Dr. River in Ado-Ekiti, Ekiti State, Nigeria. International Journal of Environmental O.Y. Titilawo for proof-reading the manuscript. Monitoring Analysis 1, 27-33. https://doi.org/ 10.11648/j.ijema.20130101.14 Egwim, E.C., Elem, R.C. & Egwuche R.U. (2011). Proximate composition, REFERENCES phytochemicalscreening and antioxidant activity of ten selected wild edible Nigerian mushrooms. American Journal of Food and Nutrition 1(2), 89-94. Adedayo, M.R. (2011). Proximate analysis on four edible mushrooms. Journal of https://doi.org/10.5251/ajfn.2011.1.2.89.94 Applied Science and Environmental Management, 15(1), 9-11. Eyabi, E.G.D. (2001). Understanding the product and process. In: Fellows P, https://doi.org/10.4314/jasem.v15i1.65666 Axtell B (eds) A handbook for setting up and running a small food business. Adeniyi, M.A. & Odeyemi, O. (2012). Comparative cultivation of mushroom Opportunities in food processing series. ACP-EU Technical Centre for (Pleurotus florida) on corncob, wood shred and old newspaper. Proceedings of Agricultural and Rural Cooperation (CTA), Wageningen, 29-50. the 25th Annual Conference of the Biotechnology Society of Nigeria 146-150.
395
J Microbiol Biotech Food Sci / Titilawo et al. 2020/21 : 10 (3) 390-397
Falandysy, J., Szymczyk, K., Ichihashi, K., Belawski, L., Gucia, M., Frankowska, Mohan, S., Campbell, N.R.C. & Willis, K. (2009). Effective population-wide A. & Yamasaki, S.I. (2001). ICP/MS and ICP/AES elemental analysis (38 public health interventions to promote sodium reduction. Analysis 181(9), 605- elements) of edible wild mushrooms growing in Poland. Food Additives and 609. https://doi.org/10.1503/cmaj.090361 Contaminants 18(16), 503-513. Mshandete, M.A. & Cuff, J. (2007). Proximate and nutrient composition of three FAO (2012). West African food composition table. Food Policy Food Nutrition types of indigenous edible wild mushrooms grown in Tanzania and their Division, FAO Rome. utilization prospects. Journal of Food, Agriculture, Nutrition and Development Feeney M.J., Dwyer, J., Hasler-Lewis, C.M., Milner, J.A., Noakes, M. Rowes, S., 7(6), 230-238. Wach, M., Beelman, R.B., Caldwell, J., Cantoma, M.T., Castlebury, L.A., Chang, Musieba, F., Okoth, S., Mibey, R.K., Wanjiku, S. & Moraa, K. (2013). Proximate S.T., Cheskin, L.J., Clemens, R., Drescher, G., Fulgoni, V.L., Haytowitz, D.B., composition, amino acids and vitamin profile of Pleurotus citrinopileatus Singer: Hubbard, V.S., Law, D., Myrdal, M.A., Minor, B., Percival, S.S., Riscuta, G., an indigenous mushroom from Kenya. America Journal of Food Technology 1-7. Schneeman, B., Thornsbury, S., Toner, C.D., Woteki, C.E. & Wu, D. (2014). https://doi.org/10.3923/ajft.2013.200.206 Mushrooms and health summit proceedings. Journal of Nutrition 144, 1128- Muthangya, M., Mshandete, A.M., Amana, M.J., Hashim, S.O. & Amelia K. 1136. (2014). Nutritional and antioxidant analysis of Pleurotus HK 37 grown on Agave Food and Nutrition Board (2011). Dietary reference intakes for calcium and sisalana saline solid waste International Journal of Research in Biochemistry vitamin D. Institute of Medicine, National Academies. Washington, DC: National and Biophysics 4(2), 5-12. Academies Press. Nakalembe, I., Kabasa, D. & Olila, D. (2009). Indigenous knowledge and usage Food and Nutrition Board (2016). Dietary reference intakes for water, potassium, of wild mushrooms in Mid-Western, Uganda. African Journal of Animal and sodium, chloride, and sulphate. Institute of Medicine, National Academies. Biomedical Science 4 (1), 63-73. Washington, DC: National Academies Press. Nakalembe, I., Kabasa, J.D. & Olila, D. (2015). Comparative nutrient Gbolagade, J., Ajayi, A., Oku, I. & Wankasi, D. (2006). Nutritive value of composition of selected wild edible mushrooms from two agro‑ecological zones, common wild edible mushrooms from Southern Nigeria. Global Journal of Uganda. SpringerPlus 4, 433. https://doi.org/10.1186/s40064-015-1188-z Biotechnology and Biochemistry 1(1), 16-21. Nasiri, F. Tarzi, B.G., Bassiri, A. & Hoseini, S. E. (2012). Comparative study on Grober, U., Schmidt, J. & Kisters, K. (2015). Magnesium in prevention and some chemical compounds of button mushrooms (Agaricus bisporus) cap and therapy. Nutrients 7, 8199-8226. https://doi.org/10.3390/nu7095388 stipe during the first to third flushes. Annals of Biological Research 3(12), 5677- Hemilä, H. (2017). Vitamin C and infections. Nutrients 9(4), 339. 5680. https://doi.org/10.3390/nu9040339 Nova Scotia Environment (2005). The drop of water calcium and magnesium. Ijioma, B.C., Ihediohanma, N.C., Onuegbu, N.C. & Okafor, D.C. (2015). Retrieved on April 4, 2019. Nutritional composition and some anti-nutritional factors of three edible http://www.szu.cz/uploads/documents/chzp/voda/pdf/hardness.pdf mushroom species in South-Eastern Nigeria. European Journal of Food Science Nwordu, M.E., Isu, R.N. & Ogbadu, G.H. (2013). Catalogue and identification of and Technology 3(2), 57-63. https://doi.org/10.13140/RG.2.2.30839.11687 some wild edible macro-fungi in Nigeria. International Journal of Food Science Januškevičius, A. Januškevičienė, G. & Andrulevičiūtė, V. (2012). Chemical 2(1), 1-15. composition and energetic values of selected vegetable species in Lithuanian Odebode, S. O. (2005). Contributions of selected non-timber forest products to supermarkets. Veterinarija Ir Zootechnika 58, 8-12. household food security in Nigeria. Journal of Food, Agriculture and Jayakumar, T., Thomas, P. A. & Geraldine P. (2009). In-vitro antioxidant Environment 3, 138-141. activities of an ethanolic extract of the oyster mushroom, Pleurotus ostreatus. Odeyemi, O. & Adeniyi, M.A. (2015). Ecology and pictorial atlas of Nigerian Innovative Food Science and Emerging Technologies 10, 228-234. mushrooms (1st ed.). Ile-Ife, Nigeria: Signet Impressions and Design Ltd. https://doi.org/10.1016/j.ifset.2008.07.002 Odeyemi, O., Adeniyi, M.A. & Odeyemi, Y. (2014). Introduction to tropical Johnsy, G., Sargunam, S. D., Dinesh, M. G. & Kaviyarasan, V. (2011). Nutritive mycology (1st ed.) Beijing, China: Universal Academic Press. value of edible wild mushrooms collected from the Western Ghats of Ohnuki, T., Alba, Y., Sakamoto, F., Kozai, N., Niizato, T. & Sasaki, Y. (2016). Kanyakumari District. Botany Research International 4(4), 69-74. Direct accumulation pathway of radioactive cesium to fruit-bodies of edible Jonathan, S. G. (2002). Studies on vegetative growth requirements and mushroom from contaminated wood logs. Scientific Report 13, 1194-1214. antimicrobial activities and antimicrobial activities of higher fungi from Nigeria. https://doi.org/10.1038/srep29866 Ph.D. Thesis, University of Ibadan, Nigeria. Okhuoya, J.A., Akpaja, E.O., Osemwegie, O.O., Oghenekaro, A.O. & Ihayere, Jonathan, S.G., Nwokolo, V.M. & Ekpo, E.N. (2013). Yield performance of C.A. (2010). Nigeria mushrooms: underutilized non-wood forest resources. Pleurotus pulmonarius (Fries.) Quelet, cultivated on different agroforest wastes Journal of Applied Science and Environmental Management 14 (1), 43-54. in Nigeria. World Rural Observations 5(1), 22-30. https://doi.org/10.4314/jasem.v14i1.56488 Kassegn, H.H. (2018). Determination of proximate composition and bioactive Okwulehie, I.C. & Ogoke, J.A. (2013). Bioactive, nutritional and heavy metal compounds of the Abyssinian purple wheat. Food Science and Technology 4, constituents of some edible mushrooms found in Abia State of Nigeria. 1421415. Kassegn, H.H. (2018). Determination of proximate composition and International Journal of Applied Microbiology and Biotechnology Research 1, 7- bioactive compounds of the Abyssinian purple wheat. Food Science and 15. https://doi.org/10.33500/ijambr.2013.01.002 Technology 4, 1421415. https://doi.org/10.1080/23311932.2017.1421415 Okwulehie, I.C., Urama, J. & Okorie, D. O. (2014). Chemical composition and Kirkland, A.E., Sario, G.L. & Holton, K.E. (2018). The role of magnesium in nutritional value of mature and young fruiting-bodies of Pleurotus pulmonarius neurological disorders. Nutrients 10(6) 730. produced on Andropogon Gayanus straw and Khaya ivorensis sawdust. Journal https://doi.org/10.3390/nu10060730 of Pharmacy and Biological Sciences 9(3), 72-77. Knochel, J.P. (1999). Phosphorus. Nutrition in health and disease (9th ed). Okwulehie, I.C. & Odunze, E. (2004). Evaluation of the myco-chemical and Baltimore: Williams and Wilkins. mineral composition of some tropical edible mushrooms. Journal of Sustainable Kowey, P.R. (2002). The role of potassium. In: Lobo, R.A., Crosignani, P.G., Agriculture and Environment 6(2), 157-162. Paoletti, R. and Bruschi, F. (eds) Women’s Health and Menopause. Medical Olaniyi, A.A. (2000). Essential medicinal chemistry. Ibadan, Nigeria: Shaneson Science Symposia Series (17). Boston, MA: Springer. Press. Kožĺšek, F. (2003). Health significance of drinking water magnesium and Olson, R.E. (1999). Water-soluble vitamins. In: Principles of Pharmacology. calcium. National Institute of public health. Retrieved April 4, 2019. Edited by Munson, P. L., Mueller, R. A. & Bresse, G. R. (Chapter 59). New http://novascotia.ca/nse/water/docs/droponwaterFAQCalciumMagnesium.pdf York: Chapman and Hall. Kumar, M. & Puri, A. (2012). A review of permissible limits of drinking water. Onuoha, C.I. & Obi-Adumanya, G.A. (2010). Proximate analysis of Pleurotus Indian Journal of Occupational and Environmental Medicine 16, 40-44. tuber-regium (Sing) grown on different substrates. Researcher 2(10), 7-11. https://doi.org/10.4103/0019-5278.99696 Osemwegie, O.O. & Okhuoya, J.A. (2009). Diversity of macrofungi in oil palm Kumari, B. & Atri, N.S. (2014). Nutritional and nutraceutical potential of wild agroforests of Edo State, Nigeria. Journal of Biological Sciences 9 (6), 584-593. edible Macrolepiotoid mushrooms of North India. International Journal of https://doi.org/10.3923/jbs.2009.584.593 Pharmacy and Pharmaceutical Sciences 6 (2), 200-204. Oyetayo, V.O. (2009). Molecular characterisation of Termitomyces species Maftoun, P., Johari, H., Soltani, M., Malik, R., Othman, N.Z. & El Enshasy, H.A. collected from Ado Ekiti and Akure, Nigeria. Nigerian Journal of Microbiology (2015). The edible mushroom Pleurotus spp.: I. biodiversity and nutritional 23(1), 1933-1938. values. International Journal of Biotechnology for Wellness Industries 4, 67-83. Patil, S.S., Ahmed, S.A., Telang, S.M. & Baig, M.M.V. (2010). The nutritional Mattila, P., Könkö, K., Eurola, M., Pihlava, J.M., Astola, J., Vahteristo, L., value of Pleurotus ostreatus (jacq.:fr.) Kumm cultivated on different Hietaniemi, L., Kumpulalainen, J., Valtonen, M. & Piironen, V. (2001). Contents lignocellulosic agro-wastes. Innovative Romanian Food Biotechnology 7, 66-76. of vitamins, mineral elements, and some phenolic compounds in cultivated Ragunathan., R. & Swaminathan, K. (2003). Nutritional status of Pleurotus sp. mushrooms. Journal of Agriculture and Food Chemistry 49, 2343-2348. grown on various agro-wastes. Food Chemistry 80, 371-375. https://doi.org/10.1021/jf001525d https://doi.org/10.1016/S0308-8146(02)00275-3 Mattila, P., Suonpää, K. & Piironen, V. (2000). Functional properties of edible Rao, A.V. & Agarwal, S. (2000). Role of antioxidant lycopene in cancer and mushrooms. Nutrition 16(7-8), 694-696. https://doi.org/10.1016/s0899- heart disease. Journal of the American College of Nutrition 19(5), 563-569. 9007(00)00341-5 https://doi.org/10.1080/07315724.2000.10718953
396
J Microbiol Biotech Food Sci / Titilawo et al. 2020/21 : 10 (3) 390-397
Rodrı´guez, R., Jime´nez, A., Ferna´ndez- Bolan˜os, J., Rafael Guille´n, R. & Heredia, A. (2006). Dietary fibre from vegetable products as a source of functional ingredients. Trends in Food Science and Technology 17, 3-15. Salano, E.M. (2013). Assessment of heavy metal pollution in soils and water at Samburu county, Kenya. MSc Thesis, Kenyatta University, Kenya. Sanmee, R., Dell, B., Lumyong, P., Izumori K. & Lumyong, S. (2003). Nutritive value of popular wild edible mushrooms from Northern Thailand. Food Chemistry 84, 527-532. Sapers, G. M., Miller, R. L., Choi, S. W. & Cooke, P. H. (1999). Structure and composition of mushrooms as affected by hydrogen peroxide wash. Journal of Food Science 64(5), 889-892. https://doi.org/10.1111/j.1365- 2621.1999.tb15934.x Shelley, E. & Geoffrey, K. (2004). Pocket nature: Fungi. London, UK: Dorling Shelley Evans and Geoffrey Kindersley Limited. Souci, S.W., Fachmann, W. & Kraut, H. (2000). Food composition and nutrition tables. 6th ed. Deutsche Forschungsanstalt f¨ur Lebensmittelchemie, Garching b. M¨unchen, Medpharm Scientific Publishing, Stuttgart, Germany. Boca Raton, London, New York, Washington, DC: CRC Press. Swaminathan, S., Seshadri, M. & Karagasapathy, A.S. (2011). Effect of tannery effluent on the zinc content of groundwater. Journal of Pharmaceutical and Biomedical Science 11, 1-3. Thatoi, H. & Sindevsachan, S.K. (2014). Diversity, nutritional composition and medicinal potential of Indian mushrooms: a review. African Journal of Biotechnology 13(4), 523-545. Titilawo, Y., Adeniji, A., Adeniyi, M. & Okoh, A. (2018). Determination of levels of some metal contaminants in the freshwater environments of Osun State, Southwest Nigeria: A risk assessment approach to predict health threat. Chemosphere 211, 834-843. https://doi.org/10.1016/j.chemosphere.2018.07.203 Usha, S. & Suguna, V. (2014). Investigation on the nutritional value of edible mushrooms viz., Auricularia, Polytricha, and Pleurotus ostreatus. Asian Journal of Science and Technology 5(8), 497-500. Vetrimurugan, E., Brindha, K., Elango, L. & Ndwandwe, O.M. (2017). Human exposure risk to heavy metals through groundwater used for drinking in an intensively irrigated river delta. Applied Water Science 7, 3267-3280. https://doi.org/10.1007/s13201-016-0472-6 Wani, H.A., Bodha, R. H. & Wani, A. H. (2010). Nutritional and medicinal importance of mushrooms. Journal of Medicinal Plants Research 4(24), 2598- 2604. Wardlaw, G.M. & Kessel, M.W. (2002). Minerals: dietary needs, absorption, transport, and excretion. In: Perspectives in nutrition, 5th ed. Mc Graw-Hill Companies Inc. Wasser, S. (2002). Medicinal mushrooms as a source of anti-tumor and immunomodulating polysaccharides. Applied Microbiology and Biotechnology 60, 258-274. doi: 10.1007/s00253-002-1076-7. WHO (2006). Reducing salt intake in populations: report of a WHO forum and technical meeting. Paris, France. Zoberi, M.H. (1973). Some edible mushrooms from Nigeria. Nigerian Field 38, 81-90.
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CAROTENOID AND ANTIOXIDANT RETENTION OF THE DEHYDRATED TOMATO PRODUCTS AFFECTED BY THEIR DIFFERENT TECHNOLOGICAL TREATMENTS
Andrea Mendelová1, Ľubomír Mendel2, Martina Fikselová3, Ján Mareček1
Address(es): doc. Ing. Andrea Mendelová, PhD. 1Department of Technology and Quality of Plant Products, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture, Tr. A. Hlinku 2, 949 76 Nitra, Slovak Republic. 2National Agricultural and Food Centre, Research Institute of Plant Production, Bratislavská cesta 122, 921 68 Piešťany, Slovak Republic. 3Department of Food Hygiene and Safety, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture, Tr. A. Hlinku 2, 949 76 Nitra, Slovak Republic.
*Corresponding author: [email protected] doi: 10.15414/jmbfs.2020.10.3.409-412
ARTICLE INFO ABSTRACT
Received 21. 8. 2019 Tomatoes are the most commonly preserved vegetables in the world. Traditional tomato products are tomato juice, puree or ketchup but Revised 27. 7. 2020 demand for dried tomatoes is increasing at the market. Drying is one of the oldest methods of preservation and often accompanies the Accepted 11. 8. 2020 degradation of nutritionally important ingredients. In order to protect nutrients from excessive oxidation during drying, the right choice Published 1. 12. 2020 of the drying temperature and the treatment of the raw material prior to drying are important elements of the manufacturing process. In our work we investigated the impact of various ways of antioxidant treatment of the raw material before drying on the stability of total carotenoids, polyphenols and antioxidant activity of the product. We used Uno Rosso F1 cultivar of tomatoes, dried by air at 70 °C, and Regular article the fruits were cut into slices of 3 mm thickness. To increase the stability of phytonutrients, we used the slice treatment prior to drying with 1% potassium bisulphite, ascorbic acid, citric and acetic acid and 5% sucrose and sodium chloride solutions. We found that the 1% solution of ascorbic acid was the most effective in protecting of total carotenoids and polyphenols. In the protection of total polyphenols, acetic acid and potassium bisulphite solutions were equally effective. The most significant increase in antioxidant activity was found to be ascorbic acid solution.
Keywords: drying, tomato, treatment
INTRODUCTION Optimizing the tomato drying process can be achieved by providing maximum drying speed and minimizing oxidative and thermal damage. The prerequisite for At the food product market, there has been an increasing demand for dried achieving the desired quality and stability of the colors is the acceleration of the tomatoes, which are used as flavoring or decorative ingredient in food (Marfil et drying process due to the reduction of the cut thickness of the dried tomatoes. It al., 2008). Dried tomatoes, tomato powder, as well as other processed products is recommended the fruit to be cut into smaller pieces before drying, eg. slices, are subject to scientific studies due to the high antioxidant activity and high quarters. This operation will provide the larger contact area of the dried product carotenoid content especially lycopene (Chang et al., 2006; Monteiro et al., with flowing air and will require less time to achieve the same level of moisture 2008; Kobori et al., 2010 ). removal (Giovanelli et al., 2002; Sanjinez-Argandoña et al., 2011). The basic preservative principle of drying is to reduce water activity, limiting In order to improve the quality of dried tomatoes, it is recommended that microbiological activity, enzymatic processes, and minimizing physical and tomatoes should be treated with calcium chloride solutions before the drying chemical changes during storage of finished products (Maskan, 2000; Krokida process (Lewicki et al., 2002; Lewicki and Michaluk, 2004), sodium chloride et al., 2001). (Sacilik et al., 2006) or sodium metabisulphite (Akanbi et al., 2002; Santos- Karam et al. (2016) report that drying is a widespread and widely used process Sanchez et al., 2012). of fruit and vegetables preserving in which the removal of water from the Latapi and Baret (2006) report that the most widely used food additive in the product minimizes the various product degradations caused either by drying industry is sulfur dioxide used in the gaseous state or as the sulfur dioxide microorganisms or by enzymes that require sufficient water content in the salt used as a solution. Santos-Sánches et al. (2012) used the solution of 1% environment. The drying conditions as well as the method of product processing sodium metabisulphite in their work. The authors report that sulfur dioxide and significantly affect the nutritional quality of the product, but also physical, sulphites are used as antioxidants to prevent degradation of ascorbic acid and textural and sensory properties. Traditionally, warm air drying techniques are the lycopene during drying, but also during storage. Muratore et al. (2008) most widely used, which may have an adverse effect on the quality of the recommend tomatoes to be treated with a 1% citric acid solution before drying. product, especially due to the ongoing physico-chemical changes that occur Abreu et al. (2011) used osmotic solutions of sugar and sodium chloride at during drying in the tissues. During drying in tomatoes, changes associated with concentrations of 5 and 10% in order to increase the proportion of soluble degradation of nutritionally important components, e.g. vitamin C and lycopene substances in the final product and to speed up the drying process. occure. Incorrect choice of drying conditions, especially low or too high The aim of this study was to evaluate the efficacy of tomato slice treatment temperature or too long drying time, can cause serious damage to the product, before drying with ascorbic, citric, acetic, potassium disulfite, sucrose and especially its taste, color and nutritional content (Doymaz, 2007; Heredia et al., sodium chloride solutions for the stability of total carotenoids, total polyphenols 2007; Cruz et al., 2012). and antioxidant activity. Color change is most often caused by degradation of carotenoids and lycopene and also by processes of non-enzymatic browning in the reaction of sugars and MATERIAL AND METHODS amino acids in the Maillard reaction process (Marfil et al., 2008). Several authors studied the optimal drying conditions (Sacilik et al., 2006; Doymaz, The experiment was performed using Uno Rosso F1 cultivar of tomatoes It is 2007; Sanjinez-Argandoña et al., 2011; Doymaz a Özdemir, 2014). medium early determinate variety, suitable for mechanized harvesting. It has
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strong growth with high fertility. The fruits are small, slightly elongated, they VIS). The total polyphenols content was calculated from calibration curve (GAE) weighs 60 - 70 grams and are resistant to cracking in more frequent rainfall and of the gallic acid at concentration 50-800 mg/L. irrigation. The fruits are particularly suitable for industrial processing. The antioxidant activity was determined by the Phosphomolybdenum method Drying was performed in the laboratory oven (MEMMERT UF 160) with the (Prieto et al., 1999). The principle of the method is based on the reduction of Mo possibility of controlling drying conditions, the drying temperature was 70 °C (VI) to Mo (V) by the reducing components of the sample and subsequent (Mendelová et al., 2014; Andrejiová and Mendelová, 2016). Drying was formation of the phosphate complex Mo (V) in acid pH with varying intensity of performed until the moisture content of the product was less than 18%. The final green color. The preparation of samples for analysis was consistent with the moisture content was monitored at (KERN MRS 120-3) moisture analyzer. The preparation of the sample for analysis of total polyphenols. The absorbance of the average drying time was 17 hours. samples was measured at JENWAY (6405 UV / VIS) spectrophotometer at 695 We dried the fruit into slices of 3 mm thickness. Before drying, we used the nm. The reducing ability of the sample was calculated from the ascorbic acid treatment of slices with antioxidant solutions. Based on recommendations such as calibration curve prepared from solutions with ascorbic acid (AAE) Latapi and Barret (2006), Hasan and El Hana (2008), Muratore et al. (2008), concentrations of 40-300 mg/L. Abreu et al. (2011), Hasturk et al. (2011), Santos-Sánchez et al. (2012), The experiment was performed in three replicates. To assess statistically Doymaz and Özdemir (2014), Azzez et al. (2017), we prepared: 1% potassium significant differences among tomato variants treated before drying, we used the metabisulphite solution, 1% ascorbic acid solution, 1% citric acid solution, 1% Tukey HSD multiple comparison test at P <0.05. acetic acid solution, 5% sucrose solution and 5% sodium chloride solution. Treatment with solutions of organic acids and potassium metabisulphite was RESULTS AND DISCUSSION performed for 5 minutes, treatment with osmotic solutions for 60 minutes. After treatment, the excess moisture of the fruits was removed on the filter paper and Several scientific works (Kerkhofs et al., 2005; Chang et al., 2006; Santos- then placed on a dryer. Sánches et al., 2012; Gümüsay et al., 2015) state that in practice the most The total carotenoid content was determined by the methodology of Hegedűsová frequent air drying causes a significant decrease in nutritional quality. In order to et al. (2016). Sample preparation for analysis consisted of homogenization at minimize losses during such drying and improve the nutritional quality of the BOSH MKN 6003 mill for 3 minutes. Carotene extraction from the homogenized final product, an important technological step is the antioxidant treatment of the samples was performed with acetone. The acetone extract obtained was shaken material prior to drying. repeatedly with petroleum ether, and the layers were phased with distilled water. Based on the measured results in total carotenoid content, total polyphenol Subsequently, the carotene-saturated petroleum ether phase was dried over content and antioxidant activity, we found that treatment of tomato fruit before anhydrous sodium sulfate, transferred quantitatively to a volumetric flask and drying showed better results in all treatments and parameters than drying without made up to the specified volume with 25 ml petroleum ether. The absorbance of pretreatment. the samples was measured spectrophotometrically at JENWAY In terms of the overall carotenoid content assessment, ascorbic acid treatment spectrophotometer (6405 UV / VIS) at 450 nm. was determined to be the most appropriate treatment method, with the highest The determination of total polyphenols was performed by the Folin-Ciocalteau increase in total carotenoid content (29.94 mg/100 g DM) compared to the method of Lachman et al. (2003). The method is based on the reaction of Folin- untreated control. This claim was also supported by the results of the Tukey HSD Ciocalteu reagent with polyphenols present in the analyzed sample to produce a assay, in which ascorbic acid treatment was statistically significant (P <0.05) the blue color product. Sample preparation for polyphenol analysis consisted of best treatment. The Tukey HSD test did not show the statistically significant homogenization and extraction in 80% ethanol at HEIDOLPH GSL 3006 shaker difference within the effect of the remaining treatments (1% potassium at 150 rpm for 24 hours. The polyphenol content was measured metabisulphite, 5% sodium chloride, 5% sucrose, 1% citric acid, 1% acetic acid) spectrophotometrically at 765 nm used JENWAY spectrophotometer (6405 UV / on total carotenoid content in dried tomato slices (Table 1).
Table 1 Means and homogeneous groups for the total carotenoid, polyphenol and antioxidant activity content at each pre-drying treatment based on multiple comparison from the Tukey HSD test Total carotenoid content Total polyphenol content Antioxidant activity Treatment (mg/100 g DM) (mg GAE/100 g DM) (mg AAE/100 g) DM control (no treatment) 126.85 ±0.29a 204.44 ±6.93a 351.37 ±16.59a citric acid 1% 130.33 ±0.42ab 218.90 ±4.22ab 388.05 ±0.29b sucrose 5% 134.79 ±0.40ab 226.35 ±5.44bc 411.22 ±10.15bc sodium chloride 5% 143.92 ±0.08ab 234.55 ±2.75c 419.04 ±6.79bc acetic acid 1% 151.42 ±2.26ab 235.36 ±4.11cd 430.21 ±5.24c potassium metabisulphite 1% 152.43 ±17,64ab 235.38 ±7.25cd 434.29 ±15.59c ascorbic acid 1% 156.79 ±19.53b 250.25 ±6.87d 540.98 ±20.47d - different letters at mean represent statistically significant differences among treatments (P <0.05)
By evaluating the total polyphenol content we found, similarly to the total to the citric acid solution (411.22 mg AAE/100 g DM) and sodium chloride carotenoid content, statistically significant (P<0.05) the highest total polyphenol (419.04 mg AAE/100 g DM). content after treatment with 1% ascorbic acid (250.25 mg GAE/100 g DM). Muratore et al. (2008) investigated the effect of drying and treatment Compared to the control sample with no treatment (204.44 mg GAE/100 g DM), temperature on changes in lycopene, β-carotene and ascorbic acid, during drying it is the highest difference (45.81 mg GAE/100 g DM). There were statistically at various temperatures as well as the effect of fruit treatment prior to drying. significant (P<0.05) different 11 test pairs. Treatment with 1% ascorbic acid was Cherry tomato fruits were treated with 1% sodium chloride solution and 1% citric not statistically significant different (P>0.05) comparing to the treatment with 1% acid prior to drying. Drying was performed at 40, 60 and 80 °C. They dried the acetic acid and 1% potassium metabisulphite (Table 1). samples to the relatively high residual moisture up to 40% and named the product Treatments with 1% citric acid solution and 5% sodium chloride solution were as semi-dry cherry tomatoes. They found that the temperature of 80 °C was the evaluated to be the less effective treatments for polyphenol protection. Treatment most suitable for the stability of lycopene and β-carotene for both untreated and with 5% sucrose solution (218.90 mg GAE/100 g DM) was not statistically treated samples. The fresh sample contained 99.8 mg/100 g DM of lycopene, and significant (P>0.05) different from the untreated control sample (204.44 mg only 58.5 mg of 100 g-1 DM was retained in 40 ° C dried sample and in the GAE/100 g DM) in total content polyphenols in dried tomato slices. sample dried at 80 °C, 76.4 mg/100 g DM. In the treated samples, the authors did As the statistically significant (P <0.05) the most effective in antioxidant not find significantly higher contents of lycopene or β-carotene compared to protection was the solution of 1% ascorbic acid again. Ascorbic acid is untreated samples. However, the positive effect of the treatment was shown in considered to be the significant antioxidant, it increased the antioxidant potential relation to ascorbic acid. In the fresh sample of tomatoes, they detected 433.5 of dried tomatoes, that can be seen on the high antioxidant activity (540.98 mg mg/100 g DM, in the untreated sample dried at 40 °C, the ascorbic acid content AAE/100 g DM). Based on the one-way ANOVA and post-hoc Tukey HSD tests, was 311.5 mg/100 g DM and in the treated sample 368.1 mg/100 g DM. it can be concluded that all 5 pre-drying treatments applied showed the Abreu et al. (2011) used 5 and 10% osmotic solutions of sucrose and sodium statistically significant (P <0.05) positive effect on antioxidant activity of dried chloride as well as various combinations prior to tomato drying. They used tomato slices (Table 1). Bonus variety, the treatment with osmotic solutions was performed for 120 Table 1 shows positive effect of the individual treatments on the antioxidant minutes, followed by drying at 65 °C for 12 hours. Treatment with osmotic activity of the dried slices. Relatively similar in antioxidant protection of dried solutions after drying resulted in the higher soluble dry matter content, especially tomato slices were solutions of potassium metabisulphite, acetic acid, sodium in solutions with 10% sucrose content. They found interesting results in the chloride and citric acid, among which we did not find any statistically significant evaluation of lycopene retention in dried samples. Sucrose solution showed the difference (P> 0.05). Statistically significant (P<0.05) the lowest effect in best results. After treatment with this solution, they detected an increase in the antioxidant protection showed the sucrose solution (388.05 mg AAE/100 g DM), lycopene content in the sample by 5.63%. The lycopene content increased from but even this solution showed no statistically different (P<0,05) results compared 446.85 μg/g DM to 472.02 μg/g DM. 10% sucrose solution and 5% sodium chloride solution were also found to be suitable solutions for treating of tomatoes
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prior to drying to improve lycopene stability. In the case of 10% sucrose solution, dehydration pre-treatment. Journal of Food Measurement and Characterization, the lycopene content after drying was 43.5% higher than in the control sample 11(4), 2247-2258. https://doi.org/10.1007/s11694-017-9609-z and 34.1% in the case of 5% sodium chloride solution. In our work, comparing to GIOVANELLI, G., ZANONI, B., LAVELLI, V., NANI, R. 2002. Water 5% solution of sucrose and 5% sodium chloride solution, we found better results sorption, drying and antioxidant properties of dried tomato products. Journal of with sodium chloride solution in relation to the stability of total carotenoids, even Food Engineering, 52(2), 135-141. http://dx.doi.org/10.1016/S0260- though it is far from being such a high retention percentage as reported by the 8774(01)00095-4 authors. In case of 5% sodium chloride solution, the total carotenoid content was GÜMÜSAY, O. A., BORAZAN, B. A., ERCAL, N., DEMIRKOL, O. 2015. higher by 13.8% compared to the control sample and 6.8% by the 5% sucrose Drying effects on the antioxidant properties of tomatoes and ginger. Food solution. Chemistry, 173, 156-162. http://dx.doi.org/10.1016/j.foodchem.2014.09.162 Hastur Sahim et al. (2011) compared the effect of different ways of drying in HASAN, N., EL HANA, A. 2008. Drying kinetics of osmotically-treated tomato slices: in the sun, in a hot air dryer at 65, 75 and 85 °C under vacuum, by tomatoes. Misr Journal of Agricultural Engineering, 25(3), 957-979. ISSN 1687- lyophilization and also by treatment before drying with 1% ascorbic acid + 1% 384. citric acid + 4% potassium carbonate solution + 2% sodium metabisulphite on the HASTURK SAHIN, F., AKTAS, T., ORAK, H., ULGER, P. 2011. Influence of stability of lycopene and the color properties of dried tomatoes. Authors found pretreatments and different drying methods on color parameters and lycopene that treatment with the prepared solution showed the positive effect on the content of dried tomato. Bulgarian Journal of Agricultural Science, 17(6), 867- retention of lycopene in dried tomatoes in all hot air drying processes as well as 881. ISSN 1310-0351. in vacuum drying and lyophilization. By evaluation of the drying in a hot-air HEGEDŰSOVÁ, A., MEZEYOVÁ, I., ANDREJIOVÁ, A. 2016. Metódy dryer as the most commonly used drying method in practice, the drying at 65 ° C stanovenia vybraných biologicky aktívnych látok. Nitra : Slovenská after the previous treatment the highest stability of lycopene was positively poľnohospodárska univerzita, 75 s. ISBN 978-80-552-1420-7. evaluated. HEREDIA, A., BARRERA, C., ANDRES, A. 2007. Drying of cherry tomato by Latapi and Barret (2006a) dried tomatoes in direct sunlight at an average daily a combination of different dehydration techniques. Comparison of kinetics and temperature of 30 °C for 9 days. In order to protect the nutritionally important other related properties. Journal of Food Engineering, 80(1), 111-118. components, they treated tomatoes before drying with 10% sodium chloride https://doi.org/10.1016/j.jfoodeng.2006.04.056 solution, 8% sodium metabisulphite solution and solution obtained by mixing of CHANG, C. H., LIN, H., CHANG, C., LIU, Y. 2006. Comparisons on the a basic solution of sodium chloride and potassium metabisulphite. They showed antioxidant properties of fresh, freeze-dried and hot-air-dried tomatoes. Journal positive effect on the stability of lycopene, not only during the drying process, of Food Engineering, 77(3), 478-485. but also during the subsequent storage of the finished products. By assessing the http://dx.doi.org/10.1016/j.jfoodeng.2005.06.061 impact on the stability of ascorbic acid, authors noted the inadequate effect of the KARAM, M. C. E., PETIT, J., ZIMMER, D., DJANTOU, E. B., SCHER, J. solutions used, as the ascorbic acid content of the dried samples decreased from 2016. Effects of drying and grinding in production of fruit and vegetable the original value of 3.37 mg/g to less than 0.4 mg/g in the treated samples. With powders: a review. Journal of Food Engineering, 188, 32-49. only sodium chloride treatment, ascorbic acid content was reduced by 97.3%, https://doi.org/10.1016/j.jfoodeng.2016.05.001 almost to zero content. KERKHOFS, N. S., LISTER, C. E., SAVAGE, G. P. 2005. Change in Colour Doymaz and Őzdemir (2014) treated tomatoes with 3% potassium carbonate and Antioxidant Content of Tomato Cultivars Following Forced-Air Drying. solution and 2% solution of oleic acid and ethanol. They used the drying Plant Foods for Human Nutrition, 60(3), 117-121. temperature of 60, 68 and 76 °C, and the tomatoes were cut in half, quarter and https://doi.org/10.1007/s11130-005-6839-8 eighth. In their conclusions they note the positive effect of the drying temperature KOBORI, C. N., HUBER, L. S., KIMURA, M., RODRIGUEZ-AMAYA, D. B. and the fruit treatment before drying on the drying rate. 2010. Teores de carotenoides em produtos de tomate. Revista Instituto Adolfo Lutz, 69(1), 78-83. ISSN 0073-9855 CONCLUSION KROKIDA, M. K., MAROULIS, Z. B., SARAVACOS, G. D. 2001. The effect of the method of drying on the colour of dehydrated products. International Based on the results, we can conclude that all pre-drying treatments used, showed Journal of Food Science + Technology, 36(1), 53-59. positive effect on the resulting nutritional quality of the dried tomato slices. In https://doi.org/10.1046/j.1365-2621.2001.00426.x assessing the effect of treatment on carotenoid stability, we found the highest KROKIDA, M. K., TSAMI, E., MAROULIS, Z. B. 1998. Kinetics on color total carotenoid content after treatment with 1% ascorbic acid solution, but based change during drying of some fruit and vegetables. Drying Technology, 16, 667- on the results of the Tukey test, this treatment was not statistically significant (P> 685. http://dx.doi.org/10.1080/07373939808917429 0.05) compared remaining treatments. By evaluating the effect of the pre-drying LACHMAN, J., PRONĚK, D., HEJTMÁNKOVÁ, A., PIVEC, V., FAITOVÁ, treatment on the stability of the total polyphenol content, treatment with 1% K. 2003. Total polyphenol and main flavonoid antioxidants in different onion ascorbic acid solution was the most effective treatment, but 1% acetic and (Allium cepa L.) varieties. Scientia Horticulturae, 30, 142-147. ISSN 1805-9333 potassium metabisulphite solutions were equally effective, with no statistically LATAPI, G., BARRETT, M. 2006. Influence of pre-drying treatments on quality significant difference (P> 0.05). The weakest effect on polyphenol stability and safety of sun-dried tomatoes. Part I: use of steam blanching, boiling brine showed the 5% sucrose solution. blanching, and dips in salt or sodium metabisulfite. Journal of Food Science, The ascorbic acid solution was also proven to be effective as pre-drying treatment 71(1), 24-31. https://doi.org/10.1111/j.1365-2621.2006.tb12401.x regarding the antioxidant activity, and contributed to increasing the antioxidant LEWICKI, P. P., LE, H. V., POMARANSKA-LAZUKA, W. 2002. Effect of pre- potential of dried tomato slices as well. Less effective than 1% ascorbic acid treatment on convective drying of tomatoes. Journal of Food Engineering, 54(2), solution appeared to be 1% acetic acid solution and 1% potassium metabisulphite 141-146. http://dx.doi.org/10.1016/S0260-8774(01)00199-6 solution as in the case of polyphenol protection, with no statistical difference. LEWICKI, P. P., MICHALUK, E. 2004. Drying of tomato pre-treated with The decreasing gradient in the antioxidant protection effect during drying calcium. Drying Technology, 22(8), 1813-1827. http://dx.doi.org/10.1081/DRT- showed solutions of sodium chloride, citric acid and sucrose, among which we 200032777 found no statistically significant (P> 0.05) difference. MARFIL, P. H. M., SANTOS, E. M., TELIS, V. R. N. 2008. Ascorbic acid degradation kinetics in tomatoes at different drying conditions. LWT- Food Acknowledgement This research was supported by KEGA 044SPU-4/2019 and Science and Technology, 41(9), 1642-1647. KEGA 017SPU-4/20019. http://dx.doi.org/10.1016/j.lwt.2007.11.003 MASKAN, M. 2000. Kinetics of colour change of kiwifruits during hot air and REFERENCES microwave drying. Journal of Food Engineering, 48(2), 169-175. http://dx.doi.org/10.1016/S0260-8774(00)00154-0 ABREU, W. C., BARCELOS, M. F. P., SILVA, E. P., BOAS, E. V. B. E. 2011. MENDELOVÁ, A., MENDEL, Ľ., FIKSELOVÁ, M., CZAKO, P. 2014. Effect Physical and chemical characteristics and lycopene retention of dried tomatoes of drying temperature on lycopene content of processed tomatoes. Potravinárstvo, subjected to different pre-treatments. Revista do Instituto Adolfo Lutz, 70(2), 168- 7(1), 141-145. ISSN 1337-0960. http://dx.doi.org/10.1007/s11101-007-9085-x. 174. ISSN 1983-3814 MONTEIRO, C. S., BALBI, M. E., MIGUEL, O. G., PENTEADO, P. T. P. S., AKANBI, C. T., ADEYEMI, R. S., OJO, A. 2006. Drying characteristics and HARACEMIV, S. M. C. 2008. Qualidade nutricionale antioxidante do tomate sorption isotherm of tomato slices. Journal of Food Engineering, 73(2), 157-163. “tipo italiano”. Alimentos e Nutrição Araraquara, 19(1), 25-31. ISSN 0103-4235 http://dx.doi.org/10.1016/j.jfoodeng.2005.01.015 MURATORE, G., RIZZO, V., LICCIARDELLO, F., MACCARONE, E. 2008. ANDREJIOVÁ, A., MENDELOVÁ, A. 2016. Vplyv diferencovanej výživy a Partial dehydration of cherry tomato at different temperature, and nutritional spracovania na obsah vybraných bioaktívnych látok v plodoch rajčiaka jedlého quality of the products. Food Chemistry, 111, 887-891. (Lycopersicon esculentum Mill.). 1. vyd. Nitra : Slovenská poľnohospodárska https://doi.org/10.1016/j.foodchem.2008.05.001 univerzita, 79 s. ISBN 978-80-552-1605-8. PRIETO, P., PINEDA, M., AQUILAR, M. 1999. Spectrophotometric AZEEZ, L., OEDEJI, A. O., ADEBISI, S. A., ADEJUMO, A. L., TIJANI, K. O. quantification of antioxidant capacity though the formation a 2017. Chemical components retention and modelling of antioxidant activity using phosphomolybdenum complex. Analytical Biochemistry, 269, 337-341. neural networks in oven dried tomato slices with and without osmotic https://doi.org/10.1006/abio.1999.4019
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J Microbiol Biotech Food Sci / Mendelová et al. 2020/21 : 10 (3) 409-412
SABLANI, S. S. 2006. Drying of fruits and vegetables: retention of nutritional/functional quality. Drying technology, 24(2), 123-135. http://dx.doi.org/10.1080/07373930600558904 SACILIK, K., KESKIN, R., ELICIN, A. K. 2006. Mathematical modeling of solar tunnel drying of thin layer organic tomato. Journal of Food Engineering, 73(3), 231-238. http://dx.doi.org/10.1016/j.jfoodeng.2005.01.025 SHI, J. X., LE MAGUER, M., KAKUDA, Y., LIPTAY, A., NIEKAMP, F. 1999. Lycopene degradation and izomerization in tomato dehydration. Food Research International, 32(1), 15-21. http://dx.doi.org/10.1016/S0963-9969(99)00059-9
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ENCAPSULATION OF POMEGRANATE SEED OIL USING W/O/W NANO-EMULSION TECHNIQUE FOLLOWED BY SPRAY DRYING AND ITS APPLICATION IN JELLY FORM
Marwa Hanafy Mahmoud*1, Fathy Mohamed Mehaya1 and Ferial Mohamed Abu-Salem1
Address(es): 1 National Research Centre, Food Technology Dept., El-Bohous St., Dokki, Cairo, Egypt.
*Corresponding author: [email protected] doi: 10.15414/jmbfs.2020.10.3.449-453
ARTICLE INFO ABSTRACT
Received 31. 3. 2020 Punicic acid, which is an ω-5 polyunsaturated fatty acid representing the main component of pomegranate seed oil (PSO) with Revised 25. 8. 2020 pharmaceutical and food usages, is susceptible to oxidation. Also, its water insoluble properties allow only few fractions to be absorbed. Accepted 23. 9. 2020 Therefore, the protection of this oil as well as bioavailability and good abortion properties are the aim of our work. Formation of water Published 1. 12. 2020 in oil in water (W/O/W) emulsion from pomegranate seed oil followed by spray-drying resulted in its transformation to nano- encapsulated oil powder whose nano-capsules had a narrow size distribution and small diameters in addition to zeta potential of about −22.9 mV. Thus, the oil-loaded system was assumed to be stable. Scanning electron microscope (SEM) measurement has confirmed the Regular article presence of surface pores which allow higher air permeability of powder, consequently a higher solubility and release was predicted. Addition of the obtained nano-encapsulated oil to jelly had a good sensory attribute and acceptability. In conclusion, pomegranate seed oil can be consumed as nano-capsules in jelly form with better water-soluble properties and it was predicted to have good absorption with expected great bioavailability.
Keywords: pomegranate seed oil, punicic acid ω-5, W/O/W nano emulsion, encapsulated oil, jelly form
INTRODUCTION including cyclodextrin inclusion, microencapsulation and mixing with edible oils or fats. Fruit processing generates a huge amount of fruit waste and by-products annually The application of encapsulation technology in functional food represents a novel worldwide which lead to serious industrial and environmental problems. So, area for researches. Nano-encapsulation of bioactive compounds could improve maximizing the reuse of fruit waste and by-products are of great demands for their dietary uptake, absorption and bio-availability of supplementary nutrients food manufactures. In this respect, fruit by-products may be used as good sources compared to ordinary sources (Mohammadi et al., 2016). of bioactive and nutritional ingredients for the production of functional foods. Since, punicic acid is extremely susceptible to oxidation and insoluble in water. Pomegranate is considered one of the tropical fruits widely grown in many Therefore, the aim of this study is to increase solubility and, accordingly, to countries (Fadavi et al., 2005). Pomegranate juice containing high amount of increase absorption and bio-availability as well as good protection of phenolic compounds such as anthocyanin, which has strong antioxidant activities pomegranate seed oil by nano-encapsulation after the formation of double (Mahmoud et al., 2017), in addition it has the potential for exploration of emulsion W/O/W and its application as a dietary supplement in the form of jelly. broader applications as good candidate for functional foods and as nutraceutical plant based products (Fouad et al., 2015). During the last few years, there are MATERIALS AND METHODS many pomegranate industrial products such as juice, jelly and jam that could produce great amounts of by-products like peels and seeds (Seeram et al., 2005). Preparation of pomegranate seed oil Seeds account for approximately 22% of the total wastes (rind plus seed) of pomegranate juice industries (FAO, 2011; Abid et al., 2017). Pomegranate seed Plant material: Pomegranate granatum fruit was purchased from local market, is rich in oil that accounts for 20% of the seed weight and contains more than Cairo, Egypt at season 2018. Pomegranate seed oil was prepared as follows; after 95% triglycerides (Lansky and Newman, 2007). This oil contains punicic acid, juice extraction, seeds were obtained then dried in air-oven at 40 oC. About 100 approximately 80% (9c,11t,13c-18:3), linoleic acid, palmitoleic acid, palmitic gm seeds were ground and extracted in duplicate with 2000 ml hexane at room acid, steric acid and arachidic acid (Hornung et al., 2002). Punicic acid is a temperature overnight. Then, the hexane was removed with a rotary evaporator at conjugated linolenic acid (18:3 n-5) that belongs to the class of polyunsaturated 40 oC under vacuum and the extract was dried to a constant weight then, it was fatty acids. Punicic acid has many health benefits due to its antioxidant and anti- stored at -18oC in the dark until being used. inflammatory properties (Jing et al., 2012). Cytotoxicity for tumor cells by pomegranate seed oil was examined in both mouse and human monocytic Characterization of pomegranate oil fatty acid profile by GC-MS leukemia cells (Suzuki et al., 2001) and it was found to be anticarcinogenic (Lansky and Newman, 2007). Also, it possesses strong antiobesity, antidiabetic, The derivatization of fatty acids was preformed according to Rozes et al., (1993). and antiproliferative effects as well as its significant effect on lipid metabolism Briefly, aliquot 20 mg of pomegranate oil was replaced into screw-capped tube (Aruna et al., 2016). These physiological effects make pomegranate seed oil of a and 1 ml of 1% sodium methoxide in methanol was added, then it was great therapeutic potential, although more researches are needed to confirm the homogenized in a vortex for 20 sec. Then, 1 mL of hexane was added which led previous findings (De-Melo et al., 2014). Since punicic acid is highly susceptible to separation of two layers, saponified (fatty acids) and unsaponified compounds; to oxidation, addition of antioxidants is very important to increase its stability. the organic layer (fatty acids) was collected, dried over anhydrous Na2SO4 and But unfortunately, addition of antioxidant was found to decrease its antitumor injected into GC-MS (Gas chromatography–mass spectrometry). activity. Also, the lake of solubility of pucinic acid in water leads to absorption of The GC-MS system (Agilent Technologies) gas chromatograph (7890B) was only a minor fraction of it. Considering these previously mentioned characters, equipped with mass spectrometer detector (5977A), Central Laboratories limited bioavailability of punicic acid is being a restricted factor that lowers its Network, National Research Centre, Cairo, Egypt. The GC was equipped with applications. Consequently, several trials were done to increase its applications HP-5MS column (30 m x 0.25 mm internal diameter and 0.25 μm film thickness). Analysis was carried out using helium as the carrier gas at a flow rate of 1.0
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ml/min, injection volume of 1 µl and the following temperature program: 50 °C strawberry essence powder. Next, 200 ml hot water was added to the powder in a for 1 min; rising at 20 °C /min to 200 °C and held for 5 min; rising at 3 °C/min to mixing bowl and the solution was adjusted with 20% citric acid solution to pH of 230 °C and held for 23 min. The injector and detector were held at 250 °C. Mass 4.3 at which point the preservative sodium benzoate (0.05%) was added. spectra were obtained by electron ionization (EI) at 70 eV and using a spectral Prepared jelly with different concentrations of the added encapsulated oil was range of m/z 20-550 and solvent delay 1.8 min. Identification of different stored in a 4 oC refrigerator and sensory evaluated to determine the more constituents was determined by comparing the spectrum fragmentation pattern preferred level of added concentration. with those stored in Wiley and NIST Mass Spectral Library data. Sensory evaluation Nano-emulsion preparation Sensory evaluation was conducted in the Food Technology Department, National W/O/W nano-emulsion was prepared by spontaneous emulsification according to Research Centre, Egypt. Twenty trained panelists aged between 25 and 40 years (Assadpour et al., 2017) with some modification. Briefly, biopolymer solution were selected to take part in the sensory panel. The panelists measured the was prepared; Arabic gum powder and maltodextrin (25 g each) were dissolved selected critical jelly attributes such as color, flavor, texture, appearance, after separately in boiling deionized water to prepare 200 ml solution. At the same taste and overall acceptability according to Mahmoud et al., (2017). All time, aqueous solution of whey protein concentrate (50 g) was prepared by evaluations were performed in a well-controlled laboratory. To determine the dispersing required amounts of whey protein (WPC) powder into 200 ml degree of like for jelly products (9 = like extremely, 5 = neither like nor deionized water. Then, all prepared solutions were gently stirred for at least 30 dislike, 1 = dislike extremely) according to Peryam and Girardot (1952). min on a magnetic stirrer, then they were mixed together and stored overnight at room temperature for full hydration of biopolymers. The emulsion was prepared Statistical analysis by mixing of biopolymer solution and Span 80 solution and then added oil as drop wise to the solution at high speed homogenization (20000 rpm) using a The data obtained from the study was statistically subjected to analysis of mechanical homogenizer (CAT, Unidrive 1000 D, M.Zipperer GmbH, Germany) variance (ANOVA) Snedecor and Cochran (1980). Data were presented as followed by prop-ultrasonic (SONICS, Vibra-cell, VCX750, USA). mean ± standard deviation (SD). The least significant difference (LSD) value was used to determine significant differences between means using ASSIST version Spray drying of W/O/W emulsion 7.7 beta. Values were considered significant at p ≤ 0.05.
The prepared emulsion solution was transformed into encapsulated powder by RESULTS AND DISCUSIONS spray drier (B-290, Buchi) equipped with a pressure air atomizing nozzle at 2.5 bar air pressure, inlet air temperature of 180 ± 5 °C, and outlet air temperature of Pomegranate seed oil Fatty acid profile by GC-Mass spectrum 90 ± 5 °C with a feed flow rate of 450 ml/h. The dried powder was collected and stored in dark bottle, air tight containers at 4°C until further analysis. Pomegranate seed is a rich source of oil with special characteristics, because it contains high percentage of punicic acid which is an omega-5 fatty acid. Results Encapsulation efficiency measurement of the present study revealed that total lipids of the Egyptian P. granatum seed was 15.38% on dry weight basis. Previous studies have indicated approximately The encapsulation efficiency was determined by calculating the difference similar results. Fadavi et al., (2006) reported that lipid content in sour between the amount of oil at the surface against encapsulated oil in capsules as pomegranate was 14.83% while Melgarejo and Artes (2000) reported that the described by Calvo et al., (2010). Briefly, 5 g of microcapsules were washed lipid content was 6.3–12.2% for sweet Spanish pomegranate. Table 1 shows the with 50 ml hexane under magnetic stirrer at 200 rpm for 60 sec. After 10 min, the fatty acid composition of oil extracted from pomegranate seed. It can be noticed mixture was filtered through filter paper (No. 41, Whatman, Maidstone, UK). that unsaturated fatty acids represent 89.73 % in pomegranate seed oil, where The powder residue was again washed twice with 5 ml of hexane. Hexane was punicic acid (-5) is the predominant fatty acid (64.91%) and punicic acid evaporated to constant weight by oven at 105 °C. The total oil content was isomers named alfa and beta eleostearic acids were represented as 11.3 and 2.86, extracted by Soxhlet extraction of microcapsules using 250 ml hexane for 5 g of respectively. Palmitic acid is the major saturated fatty acid with low percentage powder for 4 h. The encapsulation efficiency was calculated by the following (3.66 %). Dadashi et al., (2013) reported that punicic acid is the main fatty acid equation: identified by gas chromatography for the oil extracted from four Iranian commercial varieties of pomegranate seed and it was found in the range from Encapsulation efficiency (%) = (Total oil content - Surface oil content) / (Total 72.07 % to 73.31 %. The difference in punicic acid percentage between our oil content) ×100 . results and the results of Dadashi et al., (2013) may be attributed to pomegranate different varieties as well as different conditions of cultivation. Droplet size measurement Table 1 Fatty acid composition of pomegranate seed oil by GC-Mass Droplet size (particle size and zeta potential) of pomegranate nano-oil was Fatty Acid Relative % Library analyzed using a dynamic light scattering method (Zeta-sizer Nano Zs, Malvern palmatic acid Methyl ester 3.66 NIST14.L Instrument, Malvern, UK). To avoid multiple scattering, all samples were diluted by water. For microstructure analysis, a Zeiss optical microscope (Germany) was cis-Linoleic acid methyl ester 3.89 NIST14.L used and the obtained images were analyzed using Image J Software (Hosseini et Oleic acid methyl ester 5.91 W9N11.L al., 2015). stearic acid methyl ester 3.86 NIST14.L
pinucic acid methyl ester 64.91 W9N11.L Scanning electron microscopy of encapsulated powders alfa- Eleostearic acid methyl ester 11.3 W9N11.L The nano-encapsulated powder was sprinkled onto a two-sided adhesive tape and beta- Eleostearic acid methyl ester 2.86 W9N11.L then coated with a thin layer of gold. Morphological features of particles were 11-Eicosenoic acid methyl ester 0.86 NIST14.L then observed by a field emission scanning electron microscope (S- 4160 Cold Oleic acid amide 2.72 NIST14.L Field-Emission SEM, QUANTA, FEG 250, Thermo Fisher Scientific, USA) with ∑ CLA fatty acids 79.07 an accelerated voltage of 320 kV and photographed at 6000. ∑ Unsaturated fatty acid 89.73 -- Fourier transform infrared spectroscopy (FTIR) ∑ Saturated fatty acid 7.52 --
The structure analysis of the samples was examined by Fourier transform infrared Water/Oil/Water emulsion spectroscopy (FT-IR). The IR spectra of the samples were recorded by FT-IR 6000 spectrometer (JASCO, Japan). The spectrum was scanned in transmission Emulsions are one of the promising encapsulation and delivery technologies; they -1 mode from 400 to 4000 cm wavenumber. The dry samples were blended with have advantages such as controlled release and chemical stability of encapsulated KBr powder and pressed into a disk before spectrum acquisition. nutrients like minerals, vitamins, amino acids, polyphenols, bioactive compounds etc. (Ilyasoglu and El, 2019; Prichapan and Klinkesorn, 2014). Double Application in jelly form emulsions have many food applications such as reducing fat content, improving fatty acid profile, encapsulation of functional components and reducing sugar/salt Jelly powder was prepared according to (Mahmoud et al., 2017) with some content of foods by increasing taste intensity. Water-oil-water emulsions are modification. Samples were prepared with added encapsulated oil at primarily used in food applications. In this study, formation of double emulsion concentrations of 2.5, 5, 7.5 and 10%, while control sample was a commercial W/O/W by getting the oil particles wrapping the WPC molecules was made, then jelly powder. The treatments were first prepared by mixing 20 gm of gelatin it was covered by another layer of polymer to produce a bigger molecule. That powder and 75 gm of sugar with encapsulated oil, 0.1 gm of colors and 1 gm of emulsion was obtained by either WPC and span twenty to produce water in oil in
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water emulsion. This structure allows more protection of oil, as well as its and higher encapsulation efficiency (85.75%) which is in accordance with those transformation into more water soluble particles and allow addition of more results obtained by Assadpour and Jafari (2017). water-soluble additives to the emulsion (Buyukkestelli, and El, 2019).
Characterization of formulated Nano-capsules
The role of ω-polyunsaturated fatty acids and their delivery to the intestine in sufficient amount is beneficial due to its anti-inflammatory properties and accordingly, reduce the chronic diseases (Ilyasoglu and El, 2014). Therefore, characterization of the prepared biodegradable nanoparticles of pomegranate seed oil was done to predict the improvement in its stability and solubility.
Droplet size measurement
The Particle size and its distribution were measured using DLS (dynamic light scattering) method, which is the common and popular technique for determining nano droplet size distribution profile of suspension (Khazaei et al., 2014). The droplet size of the tested samples Figure 1 showed that oil-loaded polymer micelles had a narrow size distribution and small diameters (about 112 nm). That small size caused by adsorption onto the surface of a system or interface surfactant reduces the surface free energy and consequently decreases interfacial tension between the lipid matrix and the aqueous phase and leads to easier disruption of particles during homogenization to form smaller particles (Soleimanian et al., 2018). (a) Zeta potential reflects the stability of emulsion systems (Soleimanian et al., 2018). Also, Zeta potential reflects the repulsion strength between charged particles. Typically, achieving a good stability required a minimum zeta potential of ±30 mV for nano suspension electrostatically stabilized. Furthermore, ±20 mV zeta potential is sufficient in the case of the combination between electrostatic and steric forces. This is because the hydrophilic surfactant coat can improve the stability of emulsion by further hydration surface layer (Tamjidi et al., 2014). In this study, the polymer displayed a zeta potential of about −22.9 mV, hence the oil-loaded polymer system was assumed to be stable. The oil-loaded polymer micelles had a great surface area because of the entrapment of polymer molecules, which may be accountable for the enhancement of the negative charge on the micelle’s surfaces. It was reported that by relatively high surface charges, particles could reinforce each other with a strong electrostatic emulsion force, thus improved the stability of the system (Chen et al., 2012).
(b) Figure 2 SEM microphotographs of: a) nano encapsulated pomegranate seed oil & b) nano-oil particles with illustrating the structure of whey protein layer surrounded.
Characterization of encapsulated powder by FTIR
FTIR method is nondestructive technique for studying the functional groups of food matrices including oil. The FTIR spectra of pomegranate seed oil (PSO) and encapsulated PSO with whey protein/ gum Arabic/ Maltodextrin are illustrated in Figure 3. In PSO, the peak located at 3009 cm-1 corresponding to - C-H stretch which indicate to the double bond of an unsaturated fatty acid, while at 2925 and 2856 cm-1 the peaks correspond to the alkene and alkane stretch vibrations of hydrocarbon. These peaks had stronger intensity in pure oil than encapsulated ones. The peak around 990-980 cm-1 corresponds to the conjugated C-C double bonds (cis and trans) in punicic acid and their isomers. Peaks located at 3400 and 1637 cm-1 in the FTIR spectra of encapsulated oil are attributed to the O–H stretching, and asymmetric N-H (-NH3 +) of amide I. The amide I peak at 1637cm-1 referred to the α-helix of secondary structure of proteins in whey protein (Dybing and Smith, 1991). Moreover, the main ingredient of whey protein is β-lactoglobulin (apex 57%) which provide emulsifying properties for Figure 1 Zeta Size and Potential of nano encapsulated pomegranate seed oil. their amphiphilic molecules that can increase emulsion stability and generate desirable characteristics of emulsions as well as decrease the interfacial tension Encapsulated powder morphology by Scanning electron microscopy and between the hydrophilic and lipophilic phases (Sobhaninia et al., 2017). The Encapsulation efficiency peak at 1741 cm-1 represents to the carbonyl stretching group of the triacylglycerols in unsaturated fatty acids. Figure 2 reveals the SEM microphotographs of the powder containing encapsulated pomegranate seed oil with high content of ω-5 punicic fatty acid. Encapsulated powder consisted of WPC particles in the size of nano (about 112 nm) wrapped with pomegranate seed oil layer and every group (about 2-4 molecules) of nano particles covered by another layer of polymer to produce a bigger particle (ranged from 200 up to 2000 nm) as shown in Figure 2. Observations obtained from SEM confirmed the presence of surface pores obtained by emulsion matrix containing Span as a surfactant. This structure allowed great air permeability into the powder, while at the same time, much solubility and release are predictable due to higher surface area (Khazaei et al., 2014 ; Jafari et al., 2007). Also, samples became with more smooth surfaces, rare cracks, and no dents which indicated a better protection of encapsulated oil
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control
2.5 % 5%
Figure 3 FTIR spectra of pomegranate seed oil (PSO) and encapsulated PSO. 7.5% 10%
Jelly form of encapsulated oil Figure 4 Jelly form of pomegranate seed oil Nano encapsulated.
The nano-encapsulation process is an alternative technique to increase the oil Sensory evaluation stability and allow its incorporation in foods with high water content. Addition of pomegranate seed oil in the form of nano-capsules to jelly was done to improve The sensory evaluation mean scores of parameters were statistically analyzed and the bioavailability of the pomegranate seed oil. Jelly was selected as an the results are shown in Table 2. Results showed that no significant difference application because it is a dietary supplement of a delicious taste and of high was found in both of color and texture between control and nano encapsulated interest, especially for children. Figure 4 showed the jelly fortified with pomegranate seed oil powder (p > 0.05). On the other hand, sensory attributes pomegranate seed oil nano capsules with different concentration of addition, which represented in taste, flavor and after taste showed significant differences comparing to control sample. between control and samples with nano pomegranate seed oil powder. The control sample and sample with pomegranate seed oil nano powder at a level of 2.5 % had no significant difference in overall acceptability, while there was a significant difference (p < 0.05) between them and samples with 5, 7.5 and 10% pomegranate seed oil nano powder. Regarding overall acceptability, the panelists preferred control sample and the sample produced with 2.5% pomegranate seed oil nano powder. Generally, it has been observed that the higher the percentage of the added powder, the more the sweet taste and the more pronounced the flavor of the added strawberry. Also, the higher the amount of added nano-powder, the greater the turbidity and lack of transparency (Figure 4).
Table 2 Sensory evaluation mean scores of parameters Overall Treatment Color Taste Flavor After taste Texture acceptability Control 8.28a ±1.17 8.64a±0.49 8.46a±0.49 8.64a±0.49 8.1a±0.9 8.82a±0.4 T1 7.2a±0.98 7.92a±0.4 7.56ab±0.49 8.1a±0 7.92a±0.4 8.28a±0.4 T2 7.74a±0.8 6.84b±0.8 7.02b±0.98 6.66b±1.08 7.38a±1.17 7.2b±0.64 T3 7.2a±0.9 6.66b±0.8 6.66b±0.8 6.84b±1.03 7.38a±1.17 6.84c±0.49 T4 7.38a±1.17 6.3b±0.00 6.66b±0.49 6.66b±0.8 7.38a±1.17 6.48±0.4 LSD 5% 0 0.769 0.904 1.02 0 0.628 - All values are means of triplicate determinations ± standard deviation (SD). - Means withincolumn with different letters are significantly different at (P<0.05).
CONCLUSIONS Funding statement: This work was supported by the National Research Centre, Giza, Egypt (Grant No:11040110 /2016–2019 National Research Centre) as a The nano-form of pomegranate seed oil was encapsulated with whey protein, part of an ongoing project. Arabic gum and maltodextrin and Span as surfactant to improve its solubility, stability and bioavailability. The results of both zeta and electron microscope Competing interest statement: The authors declare no conflict of interest. showed that the formed emulsion was water in oil in water (W/O/W) with the formation of the two double layers surrounding the particles. Also, it showed that Originality of work : All data of this work is original data and not submitted encapsulated powder consisted of pomegranate seed oil particles in the size of before in any other journal. nano (about 112 nm) covered by a layer of polymer, and every group of nano oil Approving statement: All authors have read and approved the manuscript; and, particle covered by another layer of polymer to produce a bigger particle in that all are aware of its submission to JFST. micrometer size (about 200-2000 nm). On the other hand, the encapsulated powder of pomegranate oil had narrow size distribution and small diameters and Ethical approve: The panelists were informed about the objectives of the study which was indicated to be stable emulsion by the analysis of droplet size with and their permission in the form of written consent was obtained. The study dynamic light scattering method, in addition to be completely soluble in water. protocol was done in accordance with the guidelines stated by the Ethical Also, it had a larger surface area which might be responsible for the enhancement Committee of the National Research Centre and it was approved by the of the negative charges on the micelle’s surfaces. There were surface pores committee (16-4/2). obtained by emulsion matrix, which can allow higher air permeability into powder, while a higher solubility and release is predicted. The obtained results Additional information: No additional information is available for this paper. lead to a good characteristic of encapsulated oil with water soluble properties and accordingly, more bioavailability. Finally, formation of nano-encapsulated oil in REFERENCES the jelly shapes made it easy and attractive to the consumer as a source of dietary supplement of punicic acid with regarded to better water-soluble properties and Abid, M., Cheikhrouhou, S., Renard, C.M., Bureau, S., Cuvelier, G., Attia, H., & predicted good absorption and great bioavailability. Ayadi, M.A., (2017). Characterization of pectins extracted from pomegranate peel and their gelling properties. Food Chem., 215, 318- Acknowledgment: The work team is really thankful to the National Research 325.https://doi.org/10.1016/j.foodchem.2016.07.181 Centre that has funded the project (grantee No: 11040110 / 2016-201) to allow Aruna, P., Venkataramanamma, D., Singh, A. K., & Singh, R. P. (2016). Health for the potentiality to complete this research as a part of the project. benefits of punicic acid: a review. Comprehensive Reviews in Food Science and Food Safety, 15(1), 16-27.https://doi.org/10.1111/1541-4337.12171
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Assadpour, E., & Jafari, S. M. (2017). Spray drying of folic acid within nano- microwave method. Journal of Materials Science: Materials in Electronics,, emulsions: optimization by Taguchi approach. Drying Technology, 35(9), 1152- 1e5.https://doi.org/10.1007/s10854-016-4772-2 1160.https://doi.org/10.1080/07373937.2016.1242016 Prichapan, N., Klinkesorn, U., 2014. Factor affecting the properties of water-in- Assadpour, E., Jafari, S. M., & Maghsoudlou, Y. (2017). Evaluation of folic acid oil-inwater emulsions for encapsulation of minerals and vitamins. release from spray dried powder particles of pectin-whey protein nano-capsules. Songklanakarin J. Sci.Technol. 36 (6), 651-661. International journal of biological macromolecules, 95, 238- Peryam, D., Girardot, N. (1952). Advanced Taste-Test Method, Food 247.https://doi.org/10.1016/j.ijbiomac.2016.11.023 Engineering, 24 (7). 58-61. Buyukkestelli, H. I., & El, S. N. (2019). Development and characterization of Rozes, N., Garbay, S., Denayrolles, M., & Lonvaud‐Funel, A. (1993). A rapid double emulsion to encapsulate iron. Journal of Food Engineering, 263, 446- method for the determination of bacterial fatty acid composition. Letters in 453.https://doi.org/10.1016/j.jfoodeng.2019.07.026 Applied Microbiology, 17(3), 126-131.https://doi.org/10.1111/j.1472- Chen, B., Li, H., Ding, Y., & Suo, H. (2012). Formation and microstructural 765X.1993.tb01440.x characterization of whey protein isolate/beet pectin coacervations by laccase Seeram, N. P., Adams, L. S., Henning, S. M., Niu, Y., Zhang, Y., Nair, M. G., & catalyzed cross-linking. LWT-Food Science and Technology, 47(1), 31- Heber, D. (2005). In vitro antiproliferative, apoptotic and antioxidant activities of 38.https://doi.org/10.1016/j.lwt.2012.01.006 punicalagin, ellagic acid and a total pomegranate tannin extract are enhanced in Calvo, P., Hernández, T., Lozano, M., & González-Gómez, D. (2010). combination with other polyphenols as found in pomegranate juice. The Journal Microencapsulation of extra-virgin olive oil by spray-drying: Influence of wall of nutritional biochemistry, 16(6), 360- material and olive quality. European Journal of Lipid Science and Technology, 367.https://doi.org/10.1016/j.jnutbio.2005.01.006 112(8), 852-858.https://doi.org/10.1002/ejlt.201000059 Snedecor, G.W., Cochran, W.G., 1980. Statistical Methods, 7 th. IBIT Public. Dadashi, S., Mousazadeh, M., Emam-Djomeh, Z., & Mousavi, S. M. (2013). Co, Oxford. Pomegranate (Punica granatum L.) seed: A comparative study on biochemical Sobhaninia, M., Nasirpour, A., Shahedi, M., Golkar, A. (2017) Oil-in-water composition and oil physicochemical characteristics. International journal of emulsions stabilized by whey protein aggregates: Effect of aggregate size, pH of Advanced Biological and Biomedical Research, 1(4), 351-363. aggregation and emulsion pH. Journal of Dispersion Science and Technology, De-Melo, P.I.L., De-Carvalho, T., & Mancini-Filho, J. (2014). Pomegranate seed 38(9), 1366-1373.https://doi.org/10.1080/01932691.2016.1224719 oil (Punica granatum L.): a source of punicic acid (conjugated α-linolenic acid). J Soleimanian, Y., Goli, S. A. H., Varshosaz, J., & Sahafi, S. M. (2018). Human Nutri Food Sci, 2(1), 1-11. Formulation and characterization of novel nanostructured lipid carriers made Dybing, S. & Smith, D. (1991). Relation of chemistry and processing procedures from beeswax, propolis wax and pomegranate seed oil. Food chemistry, 244, 83- to whey protein functionality: A review. Cult. Dairy Prod. J., 26 (1), 4-12. 92.https://doi.org/10.1016/j.foodchem.2017.10.010 Fadavi A, Barzegar M, Azizi MH. (2006). Determination of fatty acidsand total Suzuki, R., Noguchi, R., Ota, T., Abe, M., Miyashita, K., & Kawada, T. (2001). lipid content in oilseed of 25 pomegranates varietiesgrown in Iran. J Food Cytotoxic effect of conjugated trienoic fatty acids on mouse tumor and human Compost Anal, 19, (6 - 7):676 - 680.https://doi.org/10.1016/j.jfca.2004.09.002 monocytic leukemia cells. Lipids, 36(5), 477-482.https://doi.org/10.1007/s11745- Fadavi, A., Barzegar, M., Azizi, M. H., & Bayat, M. (2005). Note. 001-0746-0 Physicochemical composition of ten pomegranate cultivars (Punica granatum L.) Tamjidi, F., Shahedi, M., Varshosaz, J., & Nasirpour, A. (2014). Design and grown in Iran. Food Science and Technology International, 11(2), 113- characterization of astaxanthin-loaded nanostructured lipid carriers. Innovative 119.https://doi.org/10.1177/1082013205052765 Food Science & Emerging Technologies, 26, 366- FAO, 2011. Global Food Losses and Food Waste-Extent, Causes and Prevention. 374.https://doi.org/10.1016/j.ifset.2014.06.012 UN FAO, Rome. Fouad, M. T., Moustafa, A., Hussein, L., Romeilah, R., & Gouda, M. (2015). In- vitro antioxidant and antimicrobial activities of selected fruit and vegetable juices and fermented dairy products commonly consumed in Egypt. Research journal of pharmaceutical biological and chemical sciences, 6(2), 541-550. Hornung, E., Pernstich, C., & Feussner, I. (2002). Formation of conjugated Δ11Δ13‐double bonds by Δ12‐linoleic acid (1, 4)‐acyl‐lipid‐desaturase in pomegranate seeds. European Journal of Biochemistry, 269(19), 4852- 4859.https://doi.org/10.1046/j.1432-1033.2002.03184.x Hosseini, A., Jafari, S. M., Mirzaei, H., Asghari, A., & Akhavan, S. (2015). Application of image processing to assess emulsion stability and emulsification properties of Arabic gum. Carbohydrate polymers, 126, 1- 8.https://doi.org/10.1016/j.carbpol.2015.03.020 Ilyasoglu, H., & El, S. N. (2014). Nanoencapsulation of EPA/DHA with sodium caseinate-gum arabic complex and its usage in the enrichment of fruit juice. LWT - Food Science and Technology, 56(2), 461- 468.https://doi.org/10.1016/j.lwt.2013.12.002 Jafari, S. M., He, Y., & Bhandari, B. (2007). Effectiveness of encapsulating biopolymers to produce sub-micron emulsions by high energy emulsification techniques. Food Research International, 40(7), 862- 873.https://doi.org/10.1016/j.foodres.2007.02.002 Jing, P., Ye, T., Shi, H., Sheng, Y., Slavin, M., Gao, B., Liu, L., & Yu, L., (2012). Antioxidant properties and phytochemical composition of China-grown pomegranate seeds. Food Chem., 132, 1457- 1464.https://doi.org/10.1016/j.foodchem.2011.12.002 Khazaei, K. M., Jafari, S. M., Ghorbani, M., & Kakhki, A. H. (2014). Application of maltodextrin and gum Arabic in microencapsulation of saffron petal's anthocyanins and evaluating their storage stability and color. Carbohydrate polymers, 105, 57-62.https://doi.org/10.1016/j.carbpol.2014.01.042 Lansky, E. P., & Newman, R. A. (2007). Punica granatum (pomegranate) and its potential for prevention and treatment of inflammation and cancer. Journal of ethnopharmacology, 109(2), 177-206.https://doi.org/10.1016/j.jep.2006.09.006 Maha, H. Mahmoud,, Wahba, H. M., Marwa, H. Mahmoud, and Badawy, I. H (2017). Newly Formulated Antioxidant Rich Dietary Supplement in Jelly Form for Alleviation of Liver Disease in Rats. Journal of Biological Sciences, 17 (7), 334-346.https://doi.org/10.3923/jbs.2017.334.346 Mahmoud, M. H., Seleet, F. L., & Foda, M. I. (2017). Effect of different concentration techniques in some properties of fresh and storage Pomegranate juice. Asian J Sci Res, 10(4), 290-298.https://doi.org/10.3923/ajsr.2017.290.298 Melgarejo P, Artes F. 2000. Total lipid content and fatty acidcomposition of oilseed from lesser known sweet pomegranateclones. J Sci Food Agric 80(10):1452 - 1454.https://doi.org/10.1002/1097- 0010(200008)80:10<1452::AID-JSFA665>3.0.CO;2-L Mohammadi, K., Ranjbar, M., & Targholizadeh, H. (2016). Synthesis and characterization photoluminescence properties of Au/GrO nanocomposites by
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MODELING AND OPTIMIZATION OF THE PULSED VACUUM OSMOTIC DEHYDRATION (PVOD) PROCESS OF CARROTS IN A TERNARY SOLUTION BY RESPONSE SURFACE METHODOLOGY
Md. Raihanul Haque*1, Md. Mojaffor Hosain2, Mohammad Atikur Rahman2,3, Md. Murtuza Kamal2, Shakti Chandra Mondal2, 4, A.K.M. Monjurul Islam1
Address(es): 1Department of Food Engineering and Technology, Faculty of Engineering, Hajee Mohammad Danesh Science and Technology University, Dinajpur-5200, Bangladesh. 2Department of Food Processing and Preservation, Faculty of Engineering, Hajee Mohammad Danesh Science and Technology University, Dinajpur-5200, Bangladesh. 3Department of Food Science and Nutrition, Zhejiang University, Hangzhou 310058, China. 4Center for International Education and Development, Keimyung University, Dalgubeoldaero, Dalseo-Gu Daegu, Republic of Korea.
*Corresponding author: [email protected] doi: 10.15414/jmbfs.2020.10.3.454-460
ARTICLE INFO ABSTRACT
Received 16. 1. 2020 Optimization of the pulsed vacuum osmotic dehydration (PVOD) process of carrot slices in a ternary solution was carried out using the Revised 13. 8. 2020 response surface methodology. During PVOD, the vacuum pulse was applied at the beginning of the process for 10 min at a reduced Accepted 23. 9. 2020 pressure of 500 mm of Hg throughout all the experiments. After that, the osmotic dehydration process was continued at atmospheric o Published 1. 12. 2020 pressure according to the central composite rotatable design (CCRD). The effects of temperature (35-55 C), sugar concentration (40- 50oBrix), salt concentration (5-15%) and osmosis time restored at atmospheric pressure (10-240 min) on the responses viz. water loss (WL), solute gain (SG) and colour difference (∆E) of the dehydrated samples were assessed and statistically optimized through the Regular article desirability function approach. The models obtained for water loss, solute gain and colour difference were found suitable to describe the experimental data. It was found that the time restored at atmospheric pressure has the most significant effect on the responses at the 95% confidence level. The optimum condition was found at a temperature of 50oC, sugar concentration of 45.47oBrix, salt concentration of 7.50%, and restoration time of 67.50 min. At these optimum conditions, the water loss, solute gain and colour difference were 42.61%, 10.42% and 4.38, respectively. The predicted optimum values for independent linear variables were validated by performing triplicate experiments and the simulated data were come across similar with the experimental ones.
Keywords: Drying, pulsed vacuum osmotic dehydration, response surface methodology, sugar, salt
INTRODUCTION pressure (Fante et al., 2011; Corrêa et al., 2016). This reduction of pressure promotes the removal of internal vapor or liquid from the food material while In Bangladesh, above 30-40% of fresh fruits and vegetables are spoiled due to boosting the ingress of an outermost osmotic solution through a hydrodynamic lack of proper and timely processing. In addition, the main cause of spoilage of mechanism (HDM). The HDM creates a pressure gradient because of the fresh fruits and vegetables are their high moisture content (>80%) in nature combined action of capillary flow and pressure changes which bring down on the (Karim and Hawlader, 2005; Orsat et al., 2006). Thus, the fruits and porous structures of food tissue (Corrêa et al., 2010; An et al., 2013). Several vegetables are highly perishable, requiring food preservation techniques to researchers conducted research on optimization of osmotic dehydration of carrots maintain shelf life and quality (Singh et al., 2014; Gandolfi et al., 2018). These (Rastogi and Raghavarao, 1997; Singh et al., 2008). However, despite the wide products are generally preserved by different drying techniques (Nijhuis et al., literature available only few papers reported on the optimization of PVOD of 1998). The main aim of drying is to increase the shelf life of products by carrots in a ternary solution of sugar and salt. The efficiency of dehydration reducing their water activity. It also helps to reduce bulk volume and weight of process and lycopene reserving was also improved when the ternary solution of products, decreasing storage and transportation costs (Reinert et al., 2018). sugar-salt was used (Heredia et al., 2007; Corrêa et al., 2016). However, drying is an expensive energy-intensive process and expends about 20- Optimization of any dehydration process is therefore performed to ensure best 25% of the energy used by the food industry. The energy efficiency and quality alternative solution and get an acceptable quality product from a specified set of of dried product are two important critical key parameters in food drying alternatives (Giri and Prasad, 2007). Response surface methodology (RSM) is (Kumar et al., 2014). Therefore, osmotic dehydration is one of the promising effective statistical and mathematical techniques for performing experiments, solutions for improving energy efficiency and product quality. This is because building and developing models, and seeking for optimal conditions of desirable osmotic dehydration (OD) reduces 20-30% of energy consumption during final responses. The RSM tool aids to reduce the number of experimental runs that drying of the food product. Apart from this, it improves product quality such as provide sufficient information for statistically valid results (Yadav et al., 2012). the flavor, color and the textural properties of an ultimate product by lowering Therefore, this study aimed to get the optimized process condition of PVOD of thermal damage during drying (Lenart, 1996; Shete et al., 2018). carrots in a ternary solution of sugar-salt regarding mass transfer parameters and Osmotic dehydration is a process in which water is partly removed by dipping colour differences than the fresh sample. foods, mostly fruits and vegetables in hypertonic solutions. The diffusion of water from plant tissue to the hypertonic solution is owing to osmotic pressures, MATERIAL AND METHODS which is taken place through a semipermeable membrane. Moreover, osmotic dehydration is a simple, economical process and requires a remarkably small Procurement and preparation of raw materials energy during food processing (Shi and Maguer, 2002; Sagar and Kumar, 2010). The pulsed vacuum osmotic dehydration (PVOD) is an osmotic treatment, Fresh carrots (Daucus carota L.) were collected from the local market and stored which performed at a reduced pressure during beginning of the process, at 4-5oC. Samples were washed with tap water to remove the dirt and after that accompanied by an extensive period of osmotic dehydration at atmospheric were graded by size to eliminate the variations. Then, the carrots were cut into
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the thickness of 10±0.1 mm slices by the sharp stainless steel knife. Later, the Table 1 Independent process variables and coded levels used for experimental moisture content of fresh carrots was determined by drying the samples in an design oven at 105oC for 24 h until the final weight becomes constant (AOAC, 2005). Levels Variable Name and Units The moisture content of carrots was calculated with the total weight of moisture -2 -1 0 1 2 loss of the samples after drying to the total weight of the fresh samples ratio A Temperature (oC) 35 40 45 50 55 (Rezaul et al., 2019). The initial moisture content of the carrot samples varied Sugar from 84.8% to 85.2% (wet basis). B concentration 40 42.5 45 47.5 50 (oBrix/%) Osmotic solution preparation Salt C 5 7.5 10 12.5 15 Osmotic solutions were prepared with food-grade sugar, salt and distilled water. concentration (%) The distilled water was heated above ambient temperature and the hot water D Time (min) 10 67.5 125.0 182.5 240 mixed with sugar and salt separately until predetermined concentrations attained. The concentration of osmotic solutions was checked by a digital refractometer (HI96801, HANNA Instruments, USA). Then, the ternary solutions of sugar and The center points in the CCRD design were repeated in six times to estimate the salt were provided by mixing in a magnetic stirrer according to the experimental reproducibility of the method. The experimental design of independent variables design. in uncoded forms and values of various responses are given in Table 2.
Pulsed vacuum osmotic dehydration (PVOD) Statistical data analysis
The pulsed vacuum osmotic dehydration (PVOD) experiments were conducted in Response surface methodology (RSM) was used to determine the relative stainless steel cases. The temperature of the osmotic solution was restrained by a contributions of A, B, C and D to various responses under study such as water thermocouple and controlled by a thermostatic water bath. The vacuum pressure loss (WL), solute gain (SG) and colour difference (ΔE) of osmotically dehydrated was got by a vacuum pump which settled in the vacuum dryer. The experiments samples. The second-order polynomial (SOP) model was fitted to each of the were handled with the ternary solution of sugar and salt. The samples were response variables (Yk), as mentioned in equation 4: placed in small stainless steel perforated boxes in a single layer, steeped in the osmotic solution and then wrapped with a sheet of aluminum film to prevent n nn 1 n evaporation of the osmotic solution. In addition, the fruit to solution proportion 2 Yk b k0 b ki X i b kii Xb ikij X i X j e k …………………… (4) was controlled relatively 1:45 for all experiments (Vieira et al., 2012). The i 1 i 1i 1 j i 1 vacuum pulse was kept in the first 10 min of each experiment at 500 mm of Hg (Corrêa et al., 2010; Viana et al., 2014). After the application of vacuum, the Where, Y is the response variables and x represent the coded independent dehydration process was continued at atmospheric pressure for dehydration time k i variables (i= A, B, C and D). bk0, bki, bkii, and bkij expressed are the constant, ranges from 10 to 240 min (Singh et al., 2008; Corrêa et al., 2016). Afterwards linear, quadratic and interaction regression coefficients, respectively. the osmotic process, the samples were removed from the osmotic solutions and rinsed with tap water to stop the dehydration process. The samples were dried with absorbent paper to remove free water present on the surface. The samples The statistical models and significance of the terms in the regression equation were tested by analysis of variance (ANOVA). The model adequacies were were weighed and subsequently 10g of samples were put into the petri-dish for 2 o checked by using model analysis, lack-of-fit test, and R-Squared values (R and determination of final moisture content in an oven at 105 C for 24 h (AOAC, 2 2005). adjusted R ) analysis as outlined by various researchers (Lee et al., 2000; Weng et al., 2001). The significant terms in the model were judged from the probability Determination of mass transfer parameters level (p<5%) which was calculated from the data. Co-efficient of variance (CV) is the relative dispersion of the experimental points from the model prediction. The water loss (WL) and solute gain (SG) were calculated in accordance with the Three dimensional response surfaces were generated and the Design Expert equations 1 and 2 (El-Aouar et al., 2006). software, version 7.0.3 ( Stat Ease Inc., Minneapolis, MN) was used for numerical optimization. (w X w X ) WL(%)i i f f 100 …………………………….. (1) w i Numerical optimization and validation of process conditions XXfi (wfi (1 ) w (1 ) SG100100 100 ...... (2) A numerical optimization technique was applied for simultaneous optimization of wi the responses. For this purpose, the desired goals for each independent variable
Where, wi is the initial weight of samples (g), wf is the final weight of samples at and response were chosen, which applied to either independent variables or time t (g), Xi is the initial moisture (w.b.) and Xf is the final moisture content at responses. For optimization, the goals for responses would have maximized, time t (w.b.). minimized, target, within range and none. In addition, the independent factors were kept within the experimental domain. For finding a solution, the targets Determination of colour difference (ΔE) were associated into a comprehensive composite function, D (x), called the desirability function (Myers and Montgomery, 1996), which is defined as: The colour of carrot samples was measured using Miniscan XE plus Hunter Lab Colourimeter (USA, Model 45/0-L). The colourimeter was calibrated using white D(x) (d d ...... d )1/n ……………………… (5) plates given by the manufacturers. After that, the colour of all samples was 1 2 n examined in terms of ‘L’, ‘a’, and ‘b’ after preparing a paste of the fresh and dehydrated sample. The obtained values were recorded and compared with the Where, d1, d2... dn are responses and n expressed as the total number of responses values of fresh carrot sample. The colour difference (ΔE) was calculated by the in the measure. The function D(x) considered as the desirable areas for each equation 3, given by Alam et al., (2010). response (di). The desirability is an objective function which varied from zero to one at the desired goal. The numerical optimization technique searched to a point E [(L L)2 (a a) 2 (b b)] 2 …………………… (3) that maximized the desirability function. To attain the validation of the predicted o o o process conditions, triplicate experiments were conducted adopting the conditions determined by the optimization and compared the predicted values to Where, Lo, ao, and bo represent the readings of fresh carrot sample. Therefore, the the experimentally obtained values. measured values of Lo, ao, and bo for fresh carrot samples were 54.72, 28.01, and 37.1, respectively.
Experimental design
The variables chosen for PVOD experiments were temperature (A), sugar concentration (B), salt concentration (C), and time (D). The variable levels were selected based on the previous studies described by several authors (Telis et al., 2004; Jokić et al., 2007; Singh et al., 2007). A sequential design was performed according to a central composite rotatable design (CCRD) with the aid of the software Design Expert, version 7.0.3 (Stat Ease Inc., Minneapolis, MN, 2007). The levels of the variable in coded form and actual units are presented in Table 1.
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Table 2 Central composite rotatable design (CCRD) with experimental values of various responses Run Uncoded Process variables Responses Temp. Sugar conc. Salt conc. Time (min) Water loss Solute gain Color (oC) (oBrix/%) (%) (%) (%) difference (∆E) 1 40 47.5 7.5 67.5 36.26 8.05 6.36 2 50 47.5 7.5 67.5 41.28 11.52 5.95 3 55 45.0 10.0 125.0 47.28 12.05 5.58 4(C) 45 45.0 10.0 125.0 45.15 10.91 4.09 5(C) 45 45.0 10.0 125.0 46.34 12.95 5.15 6 45 45.0 10.0 10.0 29.82 5.10 3.97 7 50 42.5 7.5 182.5 41.92 11.79 7.99 8 40 42.5 12.5 67.5 31.03 10.58 8.90 9(C) 45 45.0 10.0 125.0 43.0 12.13 6.02 10 50 47.5 12.5 182.5 43.49 16.21 6.52 11 45 45.0 10.0 240.0 46.60 12.24 5.40 12 40 47.5 12.5 182.5 51.41 12.02 6.14 13 40 42.5 12.5 182.5 47.02 11.96 7.97 14 40 47.5 7.5 182.5 51.56 11.37 6.43 15 50 42.5 7.5 67.5 41.28 8.25 4.60 16 40 47.5 12.5 67.5 39.75 6.95 7.44 17 40 42.5 7.5 67.5 31.40 8.10 4.58 18 45 45.0 15.0 125.0 46.31 10.40 5.14 19 35 45.0 10.0 125.0 50.64 9.96 5.75 20(C) 45 45.0 10.0 125.0 46.04 11.62 5.27 21 45 40.0 10.0 125.0 41.88 9.20 8.70 22 50 42.5 12.5 182.5 41.81 11.98 5.80 23 45 45.0 5.0 125.0 41.89 10.78 5.88 24 45 50.0 10.0 125.0 49.57 11.59 8.44 25 40 42.5 7.5 182.5 45.59 10.99 8.11 26 50 47.5 12.5 67.5 43.82 10.94 5.44 27(C) 45 45.0 10.0 125.0 46.91 12.27 5.05 28 50 47.5 7.5 182.5 45.70 16.10 7.18 29(C) 45 45.0 10.0 125.0 45.68 12.83 5.30 30 50 42.5 12.5 67.5 41.54 12.51 6.34 * C=Center point
RESULTS AND DISCUSSION Where, WL= water loss (%), SG= solute gain (%), ∆E= colour difference, A= temperature (oC), B= sugar concentration (oBrix/%), C= salt concentration (%), Fitting models and D= time (min).
The design of experiments with levels of four independent variables and responses at each combination are analyzed and presented in Table 2. The values of responses were studied in the average of two replications. The second order polynomial eq. (4) was fitted to the experimental values and tested for adequacies by analysis of variance (ANOVA) are given in Table 3. The low probability value of the model for all responses (WL, SG and colour difference) also explains that the models were significant at a 95% confidence level. The ANOVA showed that the Lack-of-fit test was non-significant (p>0.05) for all responses and confirms the applicability of the statistical method used. The R2 values estimated by a least square technique for WL, SG and colour difference (∆E) were 0.94, 0.88 and 0.82, respectively, which measured a good fit of the models to the data. It also exhibits a good correlation between the actual values and predicted values Figure 1 The fitted line plot signifying the closeness between actual values and as shown in Fig. 1 (A-C). The values of adjusted-R2 were 0.88, 0.76 and 0.67 for predicted values for (A) water loss (WL) (B) solute gain (SG) and (C) colour WL, SG, and colour difference, respectively found close to the coefficient of difference (∆E) determination (R2) and suggesting that non-significant terms were not included in the model. The coefficient of variation (CV) is a measure of reproducibility of the model and shows the reliability of the experiments. A model can be regarded fairly reproducible if its CV is not greater than 10% (Shishir et al., 2016). The low variation in coefficient value for WL (4.39%) and SG (10.0%) depicted that the models of these responses were highly reliable while the higher variation in CV (12.81%) was attained for colour difference.
WL 45.09 1.96B 3.99D 3.26AD 2.12D2 (R2=0.88)...……………. (6)
2 SG 11.61 0.98A 0.49B 1.66D 0.84AB 0.69BD 0.62D (R2=0.81) …………….. (7)
E 5.45 0.39D 0.62CD 0.92B2 (R2=0.63) …………….. (8)
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Table 3 ANOVA showing the effects of the variables on water loss (WL), solute gain (SG) and colour difference (∆E) for PVOD of carrots
Source DF Water loss (WL) Solute gain (SG) Colour difference (ΔE) β Sum of Mean p-value β Sum of Mean p-value β Sum of Mean p-value square Square square Square square Square Model 14 45.09 827.87 59.13 0.0001* 11.61 134.68 9.62 0.0002* 5.45 34.01 6.80 0.0018* Temp. (A) 1 4.16E-3 4.167E-4 4.17E-4 0.9916 0.98 22.93 22.93 0.0006* -0.27 1.55 1.73 0.1369 Sugar Conc. 1 1.96 92.28 92.28 0.0001* 0.49 5.78 5.78 0.0472* -0.14 0.47 0.47 0.4009 (B) Salt Conc. 1 0.57 7.84 7.84 0.1619 0.26 1.61 1.61 0.2715 0.078 0.14 0.14 0.6364 (C) Time (D) 1 3.99 381.60 381.60 0.0001* 1.66 66.0 66.0 0.0001* 0.39 3.67 3.67 0.0285* AB 1 -1.01 16.40 16.40 0.0504 0.84 11.36 11.36 0.0084* 0.22 0.78 0.78 0.2795 AC 1 -0.24 0.96 0.96 0.6142 0.061 0.060 0.060 0.8286 -0.41 2.71 2.71 0.0548 AD 1 -3.26 169.91 169.91 0.0001* 0.013 2.5E-03 2.5E-03 0.9647 0.24 0.89 0.89 0.2495 BC 1 0.15 0.38 0.38 0.7511 -0.55 4.86 4.86 0.0660 -0.26 1.05 1.05 0.2136 BD 1 -2.5E-3 1.0E-04 1.0E-4 0.9959 0.69 7.51 7.51 0.0263* -0.27 1.19 1.19 0.1874 CD 1 -0.43 3.03 3.03 0.3751 -0.2 0.62 0.62 0.4911 -0.62 6.14 6.14 0.0062* A2 1 0.52 7.27 7.27 0.1769 -0.1 0.30 0.30 0.6295 0.21 1.04 1.04 0.2174 B2 1 -0.29 2.37 2.37 0.4316 -0.26 1.81 1.81 0.2447 0.90 24.40 24.40 0.0001* C2 1 -0.7 13.44 13.44 0.0733 -0.21 1.19 1.19 0.3421 0.17 0.87 0.87 0.2563 D2 1 -2.12 129.46 129.46 0.0001* -0.62 13.0 13.0 0.0055* -0.03 0.022 0.022 0.8553 Lack of fit 10 44.96 4.50 0.1737** 15.62 1.56 0.1453** 7.46 0.75 0.2416** R2 0.94 0.88 0.82 Adjusted- R2 0.88 0.76 0.67 C.V. % 4.39 10.0 12.81 Std. dev. 1.90 1.11 0.78 * Significant at 5% level of significance (p<0.05); β= coefficient of estimate; DF= Degree of freedom ** Non-significant at 95% confidence level (p>0.05)
Influence of process variables on the responses
The main and interactive effect of different independent variables such as temperature (A), sugar concentration (B), salt concentration (C) and restoration time at atmospheric pressure (D) were observed on the responses. Following sections describe in details about the influences of these independent variables on the responses.
Effect on water loss (WL)
The results of water loss (WL) of PVOD of carrot slices are presented in Table 2. The WL during PVOD of carrot slices was ranged from 29.82 to 51.56%. The process parameters, sugar concentration and time restored at atmospheric pressure have the positive and significant effect on water loss (p<0.05). For water loss, the magnitude of β value (Table 3) shows that the restoration time at atmospheric pressure after vacuum pulse application has the maximum positive effect on water loss followed by sugar concentration. The positive signs of linear Figure 2 Response surface plots showing the effects of process parameters on independent variables revealed that water loss increased with the increase of water loss (WL); (A) 2D contour plot (B) 3D contour plot in relation to solution restoration time and sugar concentration. Similar trends for water loss were temperature and time restored at atmospheric pressure reported by Lewicki and Lukaszuk (2000) for apple, Azoubel and Murr (2004), and An et al. (2013) for cherry tomatoes. Water loss increased with an To visualize the combined effect of different independent variables on WL, increase in the sugar concentration since the solute concentration gradient response surface 2D and 3D contour plots were generated for the fitted model as increased between the solution and the food, with the consequent increase in shown in Fig. 2 (A-B). Only the interaction term of solution temperature and pressure (Chafer et al., 2003; Viana et al., 2014). In addition, the ternary process time restored at atmospheric pressure (AD) has the significant influence solution of different concentrations was used during PVOD of carrot slices which on WL (Table 3) at the 95% confidence level. Increase in time restored at contains salt may increase the driving force of the process for the reduction in atmospheric pressure (125-182.5 min) with a combination solution temperature water content thus leading to the increase in WL (Hamledari et al., 2012). (47.5- 50oC) resulted in the reduction of water loss. This result was corroborated However, the quadratic square terms of the time restored at atmospheric pressure with the findings of Ito et al. (2007), Corrêa et al. (2010) and Ferrari et al. (D2) has the significant and negative effect on WL, whereas the quadratic terms (2011) for mango slices, guavas and melon, respectively. The samples showed of other independent variables were non-significant at p<0.05(Table 3). greater water loss from the vegetable tissues during first 120 min when restored at atmospheric pressure is because of the application of vacuum conditions. After that, a gradual stabilization of WL was observed at the end of the osmotic process. The increase of WL during the first period restored at atmospheric pressure can be explained by the hydrodynamic mechanism that was applied at the beginning of the process. With the application of pulsed vacuum conditions, the occluded gas in the intercellular spaces of the food tissues are removed and when restored at atmospheric pressure, the pores of the food material are filled by the osmotic solution since making the mass transfer easier (Deng and Zhao, 2008; Medina-Torres et al., 2008; Mundada et al., 2010). Increasing temperature above 47oC caused decreasing in the carrot samples WL. This can be related to the shorter vacuum pulse time which released the internal gas partially presence in the food spores. Hence, when the viscosity of the osmotic solution reduces as the temperature raises resultant in lessening water and mass loss because of the lack of free volume for impregnation (Torres et al., 2007).
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Effect on solute gain (SG) Effect on colour difference (∆E)
Table 2 showed that solute gain (SG) ranges from 5.10 to 16.21%. The p and β The colour differences (∆E) of carrot slices ranged from 3.97 to 8.90 (Table 2), values in Table 3 revealed that the independent process variable of temperature, which illustrated that the colour differences between lowest and highest value sugar concentration and osmosis time restored at atmospheric pressure exhibited was 76.61% after PVOD of the carrot slices at different experimental the positive and significant effect on SG at a 5% level of significance. The conditions. The co-efficient of estimate β values indicated that the time (β=0.39) relative magnitude of β value illustrates that the osmotic restoration time at restored at atmospheric pressure has a positive effects on the colour parameters atmospheric pressure was the most significant factor on the solute gain followed whereas other linear process variables have no significant effects at the 5% level by solution temperature and sugar concentration on SG (Table 3). These results of significance (Table 3). It implies that with the increase of restoration time are under the findings observed by Ferrari and Hubinger (2008), Sutar and leads to the colour degradation of the dehydrated product. Similar behavior of Prasad (2011) and Zapata et al. (2016) in melon cubes, carrots and cape changes in colour parameters were also reported by Singh et al. (2010), Moreira gooseberry, respectively. This outcome revealed that SG increased with the et al. (2011) and Najafi et al. (2014) for carrots, chestnuts and red pitaya, increase of restoration time at atmospheric pressure after the vacuum pulse respectively. The carrot contains a high content of carotenoid pigments in which conditions were applied. The higher the gain in the solute is because of the β-carotene present usually. This β-carotene may be oxidized with the increase of application of vacuum pulse at the first minutes of the process which provoke the process time and lower water activity, thus may cause in carotene bleaching opening of food pores and the exit of entrapped air. Since, when restored at which contributes to changes or losses of the colour of osmotically dehydrated atmospheric pressure for further osmotic dehydration (OD) for the greater time, carrot slices (Kidmose et al., 2002). However, the quadratic square terms of such pores are filled with the osmotic solution thus may affect the foods to solute sugar concentration has the significant effects while other variables have no gain (Huayamave and Cornejo, 2005; Zapata et al., 2016). However, only the effects on colour difference (p<0.05). quadratic square terms of time has the negative and significant effect (p<0.05) on The 2D and 3D contour response surface plots are generated for the fitted model SG while other process variables has the non-significant effect on solute gain. by keeping two other independent variables at the center points to visualize the The negative coefficient of the quadratic term of time suggested that an excessive combined effect on colour difference (Fig 4A-B). The interaction of the “salt increase in the levels of this variable resultant in the significant decrease in SG. concentration and time” has the negative and significant effect on the colour parameters (Table 3) at the 95% confidence level. The combination of a fall-off process time (125 to 67.50 min) at atmospheric pressure and with the salt concentration ranged from 7.50 to 10% in ternary solutions, thus may cause a lowering in the colour differences of the dehydrated product (Fig. 4A-B). In some vegetables such as carrots, accumulating high amounts of carotenoids in its roots are mainly prevailed by carotene pigments, which are combined with the carbon and hydrogen atoms resulting in low polar compounds. These carotene pigments of vegetables are degraded because of the oxidative browning reactions, these reactions can be limited by hindering the access of oxygen into the intercellular spaces through the application of vacuum conditions. With the vacuum pulse application, the entrapped gases occluded in the intercellular spaces of the food tissues are removed and the food spores are filled by the osmotic solution. Hence, it may reduce the availability of oxygen for oxidative reactions during PVOD and for shorter osmosis time at atmospheric pressure. So, it appears to less increase in colour difference of the dehydrated product (Talens et al., 2002; Chafer et al., 2003). Apart from this, an increasing in salt concentration over 12.50% in the ternary osmotic solutions with the increase of time suggesting that the difference in the colour parameters was decreased more than the fresh sample (Fig. 4A- B). The chromoplasts in a carrot root contain mainly large carotenoid crystals which deposited in the plant plastids. These carotenoids containing plastids are hydrophobic and degraded easily from the food matrix during dehydration and biosynthesis of that plastids may provoke the over formation of carotenoid pigments. Since, the addition of salts in ternary osmotic solution may reduce the rate of degradation of plastids, which neutralizes the negative polar surface charge of lipids and protein of plastids by the positive ions of salts (Schweiggert et al., 2012; Viana et al., 2014). Therefore, the colour parameters of the Figure 3 Response surface plots showing the effects of process parameters on dehydrated sample were decreased to a lesser extent than the fresh sample. solute gain (SG); (A-B) 2D and 3D contour plots in relation to solution temperature and sugar concentration; (C-D) 2D and 3D contour plots in relation to sugar concentration and restoration time at atmospheric pressure
To get the better understanding of the variables and their interactions on SG, the response surface contour plots are depicted in Fig. 3 (A-D) for SG during PVOD of carrots. The interaction effect of solution temperature and sugar concentration (AB) was the positive and statistically significant effect on SG at the 5% level of significance. The results showed that the SG increased with the increasing solution temperature from 40 to 50oC and sugar concentration from 42.5 to 47.5oBrix (Fig. 3A-B). The higher solution temperature favors the swelling and plasticization of the cell membrane, which make the food tissues easier to transfer water and entrance of solids. The viscosity of the osmotic solution also decreased with an increase in solution temperature resulting lower external resistance to mass transfer (Uddin et al., 2004; Lombard et al., 2008). However, the increase in osmotic solution concentration also contributed to the less increase in solute gain while used a ternary solution of sugar-salt regarding to a Figure 4 Response surface plots showing the effects of process parameters on binary solution of sodium chloride. The greater size of sugar molecules in colour difference (∆E); (A) 2D contour plot (B) 3D contour plot in relation to salt comparison with that of salt molecules could hinder their entering the cells thus concentration and time restored at atmospheric pressure lowering in the solute uptake (Heredia et al., 2007; Ferrari et al., 2011; Corrêa et al., 2016). The interaction between sugar concentration and time restored at Optimization of the process variables by the desirability function approach atmospheric pressure (BD) was significant at the 95% confidence level. The findings showed that the sugar concentration was more effective for SG at higher The numerical optimization of the pulsed vacuum osmotic dehydration (PVOD) restoration time of 182.50 min for atmospheric pressure osmotic process (Fig. of carrot slices was performed using the desirability function technique for the 3C-D). This could be attributed to the rupture of cell wall above 150 min developed models. The main criteria for constraints were to maximize WL, and resulting in higher solid incorporation. The PVOD process is accompanied with minimize SG and colour difference of the dehydrated product. However, the the hydrodynamic mechanism (HDM) which deformed food matrix by the independent variables such as temperature, sugar concentration, salt expansion and compression of the gas occluded into the food porous structure. concentration and time restored at atmospheric pressure were set in the Therefore, it alters the cell wall resistance and influences the final solid uptake experimental range for the optimization of the process variables. The optimized throughout osmotic process (Fito et al., 1996; Castelló et al., 2010). values of solution temperature, sugar concentration, salt concentration and
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restoration time found were 50oC, 45.47oBrix, 7.50% and 67.50 min, CORRÊA, J. L. G., ERNESTO, D. B., de MENDONÇA, K. S. 2016. Pulsed respectively. Therefore, the predicted water loss, solute gain, and colour vacuum osmotic dehydration of tomatoes: sodium incorporation reduction and difference values attributed under these optimum conditions were 42.61%, kinetics modeling. LWT - Food Science and Technology, 71, 17–24. 10.42% and 4.38, respectively. https://doi.org/10.1016/j.lwt.2016.01.046 DENG, Y., ZHAO, Y. 2008. Effects of pulsed-vacuum and ultrasound on the Table 4 Predicted and experimental values of the responses at optimum osmodehydration kinetics and microstructure of apples (Fuji). Journal of Food conditions for pulsed vacuum osmotic dehydration (PVOD) of carrot slices Engineering, 85(1), 84–93. https://doi.org/10.1016/j.jfoodeng.2007.07.016 Response Predicted value Experimental ±SEM EL-AOUAR, Â. A., AZOUBEL, P. M., BARBOSA, J. L., MURR, F. E. X. 2006. Influence of the osmotic agent on the osmotic dehydration of papaya (Carica Water Loss (%) 42.61 43.26 ± 0.38 papaya L.). Journal of Food Engineering, 75(2), 267–274. Solute gain (%) 10.42 10.51±0.18 https://doi.org/10.1016/j.jfoodeng.2005.04.016 FANTE, C., CORRÊA, J., NATIVIDADE, M., LIMA, J., LIMA, L. 2011. Colour difference 4.38 4.96 ± 0.26 Drying of plums (Prunus sp, c.v Gulfblaze) treated with KCl in the field and (∆E) subjected to pulsed vacuum osmotic dehydration. Internation Journal of Food Science and Technology, 46(5), 1080-1085. https://doi.org/10.1111/j.1365- * SEM= Standard error of the mean 2621.2011.02619.x FERRARI, C.C., ARBALLO, J.R., MASCHERONI, R.H., HUBINGER, M. D. Validation of the models 2011. Modelling of mass transfer and texture evaluation during osmotic dehydration of melon under vacuum. International Journal of Food Science and The optimized results were validated by conducting triplicate experiments under Technology, 46(2), 436–443. https://doi.org/10.1111/j.1365-2621.2010.02510.x the recommended optimum condition with a slight alteration in sugar FERRARI,C.C., HUBINGER, M. D. 2008. Evaluation of the mechanical concentration by 45oBrix and restoration time by 68 min in substitution of o properties and diffusion coefficients of osmodehydrated melon cubes. 45.47 Brix and 67.50 min. The predicted values of respective responses and International Journal of Food Science and Technology, 43(11), 2065–2074. experimental values are shown in Table 4. The experimental values were found https://doi.org/10.1111/j.1365-2621.2008.01824.x decent with the predicted values which satisfy the predicted response surface FITO, P., ANDRÉS, A., CHIRALT, A., PARDO, P. 1996. Coupling of model. hydrodynamic mechanism and deformation-relaxation phenomena during vacuum treatments in solid porous food-liquid systems. Journal of Food CONCLUSIONS Engineering, 27(3), 229-240. https://doi.org/10.1016/0260-8774(95)00005-4 GIRI, S.K., PARASAD, S. 2007. Optimization of microwave-vacuum drying of Response surface methodology (RSM) showed that the models obtained were button mushrooms using response surface methodology. Drying Technology, suitable to describe the experimental data of water loss (WL), solute gain (SG) 25(5), 901-911. https://doi.org/10.1080/07373930701370407 and colour difference (∆E). The process time restored at atmospheric pressure has HAMLEDARI, A., BASSIRI, A., GHIASSI T. B., BAMENI. M. M. 2012. the most pronounced effect on the mass transfer parameters and the colour Pulsed vacuum osmotic dehydration of garlic bulbs followed by microwave parameters of the osmotically dehydrated carrot slices (p<0.05). The mass drying. Journal of Food Biosciences and Technology, 2(2), 41–56. transfer rates were improved due to the expulsion of occludes gases during HEREDIA, A., BARRERA, C., ANDRE, A. 2007. Drying of cherry tomato by a PVOD of carrot slices, which induces the exchange of the water and solute in the combination of different dehydration techniques. Comparison of kinetics and food matrix. The degradation of colour parameters of the dehydrated samples was other related properties. Journal of Food Engineering, 80(1),111-118. increased with the increase of the dehydration time. The long duration of the OD https://doi.org/10.1016/j.jfoodeng.2006.04.056 could be resulted in the deteriorating of the carotenoid pigments for oxidative HUAYAMAVE, E.C., CORNEJO, F. 2005. Influence of vacuum pressures on browning reactions. For the fitted models, the optimum conditions were found to o o mass transfer during osmotic dehydration of mango. Revista Tecnológica be 50 C, 45.47 Brix, 7.50% and 67.50 min of solution temperature, sugar ESPOL, 18(1), 141-145. concentration, salt concentration and osmosis time at atmospheric pressure, ITO, A. P., TONON, R. V., PARK, K. J., HUBINGER, M. D. 2007. Influence of respectively. The various experimental response values were found closeness to process conditions on the mass transfer kinetics of pulsed vacuum osmotically the predicted values. The process variables of PVOD process could be optimized dehydrated mango slices. Drying Technology, 25(10), 1769–1777. by using RSM that shorten the number of experimental runs. Therefore, PVOD https://doi.org/10.1080/07373930701593263 pretreatment could remove a large portion of moisture at low temperature. This JOKIĆ, A., GYURA, J., LEVIĆ, L., ZAVARGÓ, Z. 2007. Osmotic dehydration may shorten the total time of final drying, which in turns reduce the loss of the of sugar beet in combined aqueous solutions of sucrose and sodium chloride. thermal sensitive nutrition and sensory quality. Journal of Food Engineering, 78(1), 47–51. https://doi.org/10.1016/j.jfoodeng.2005.09.003 REFERENCES KARIM, M. A., HAWLADER, M. N. A. 2005. Mathematical modelling and experimental investigation of tropical fruits drying. International Journal of Heat ALAM, M. S., AMARJIT, S., SAWHNEY, B. K. 2010. Response surface and Mass Transfer, 48(23–24), 4914–4925. optimization of osmotic dehydration process for aonla slices. Journal of Food https://doi.org/10.1016/j.ijheatmasstransfer.2005.04.035 Science and Technology, 47(1), 47–54. https://doi.org/10.1007/s13197-010- KIDMOSE, U., EDELENBOS,M., NØRBÆK, R., CHRISTENSEN, L. P. 2002. 0014-4 Colour stability in vegetables. In D. B. MacDougall (Ed.), Colour in food: AN, K., LI, H., ZHAO, D., DING, S., TAO, H., WANG, Z. 2013. Effect of Improving quality (pp. 179-232). CRC Press Publishing Limited, Osmotic Dehydration with Pulsed Vacuum on Hot-Air Drying Kinetics and Cambridge. http://dx.doi.org/10.1533/9781855736672.2.179 Quality Attributes of Cherry Tomatoes. Drying Technology, 31(6),698–706. KUMAR, C., KARIM, M. A., JOARDDER, M. U. H. 2014. Intermittent drying https://doi.org/10.1080/07373937.2012.755192 of food products: a critical review. Journal of Food Engineering, 121(1), 48–57. AOAC. 2005. Official methods of analysis of AOAC International. Arlington, Va: https://doi.org/10.1016/j.jfoodeng.2013.08.014 AOAC International. LEE, J., YE, L., LANDEN, W. O., EITENMILLER, R. R. 2000. Optimization of AZOUBEL, P. M., MURR, F.E.X. 2004. Mass transfer kinetics of osmotic an extraction procedure for the quantification of vitamin e in tomato and broccoli dehydration of cherry tomato. Journal of Food Engineering, 61(3), 291–295. using response surface methodology. Journal of Food Composition and Analysis, https://doi.org/10.1016/S0260-8774(03)00132-8 13(1), 45–57. https://doi.org/10.1006/jfca.1999.0845 CASTELLÓ, M. L., FITO, P. J., CHIRALT, A. 2010. Changes in respiration rate LENART, A. 1996. Osmo-convective drying of fruits and vegetables: technology and physical properties of strawberries due to osmotic dehydration and storage. and application. Drying Technology, 14(2), 391–413. Journal of Food Engineering, 97(1), 64–71. https://doi.org/10.1080/07373939608917104 https://doi.org/10.1016/j.jfoodeng.2009.09.016 LEWICKI, P. P., LUKASZUK, A. 2000. Effect of osmotic dewatering on CHAFER, M., GONZALEZ-MARTINEZ, C. FERNANDEZ, B., PEREZ, L., rheological properties of apple subjected to convective drying. Journal of Food CHIRALT, A. 2003. Effect of blanching and vacuum pulse application on Engineering, 45(3), 119–126. https://doi.org/10.1016/S0260-8774(00)00025-X osmotic dehydration of pear. Food Science and Technology International, LOMBARD, G. E., OLIVEIRA, J. C., FITO, P., ANDRÉS, A. 2008. Osmotic 9(5),321-328. https://doi.org/10.1177/1082013203039253 dehydration of pineapple as a pre-treatment for further drying. Journal of Food CORRÊA, J. L. G., VIANA, A. D., MENDONÇA, K. S., JUSTUS, A. 2016. Engineering, 85(2), 277–284. https://doi.org/10.1016/j.jfoodeng.2007.07.009 Optimization of Pulsed Vacuum Osmotic Dehydration of Sliced Tomato. In: MEDINA-TORRES, L.,GALLEGOS-INFANTE, J. A.,GONZALEZ-LAREDO, Delgado J., Barbosa de Lima A. (eds), Drying and Energy Technologies (Vol. R. F., ROCHA-GUZMAN, N. E. 2008. Drying kinetics of nopal (Opuntia ficus- 63:207-228). Switzerland:Springer International Publishing. indica) using three different methods and their effect on their mechanical https://doi.org/10.1007/978-3-319-19767-8_11 properties. LWT - Food Science and Technology, 41(7), 1183–1188. CORRÊA, J. L. G., PEREIRA, L. M., VIEIRA, G. S., HUBINGER, M. D. 2010. https://doi.org/10.1016/j.lwt.2007.07.016 Mass transfer kinetics of pulsed vacuum osmotic dehydration of guavas. Journal MOREIRA, R., CHENLO, F., CHAGURI, L., VÁZQUEZ, G. 2011. Air drying of Food Engineering, 96(4), 498–504. and colour characteristics of chestnuts pre-submitted to osmotic dehydration with https://doi.org/10.1016/j.jfoodeng.2009.08.032
459
J Microbiol Biotech Food Sci / Haque et al. 2020 : 10 (3) 424-430
sodium chloride. Food and Bioproducts Processing, 89(2), 109–115. https://doi.org/10.1016/j.foodchem.2006.01.020 https://doi.org/10.1016/j.fbp.2010.03.013 UDDIN, M. B., AINSWORTH, P., İBANOĞLU, Ş. 2004. Evaluation of mass MUNDADA, M., SINGH, B.,MASKE, S. 2010. Optimisation of processing exchange during osmotic dehydration of carrots using response surface variables affecting the osmotic dehydration of pomegranate arils. International methodology. Journal of Food Engineering, 65(4), 473–477. Journal of Food Science & Technology, 45(8), 1732– https://doi.org/10.1016/j.jfoodeng.2004.02.007 1738. https://doi.org/10.1111/j.1365-2621.2010.02328.x VIANA, A. D., CORRÊA, J. L. G., JUSTUS, A. 2014. Optimisation of the MYERS, R. H., MONTGOMERY, D. C.1995. Response Surface Methodology: pulsed vacuum osmotic dehydration of cladodes of fodder palm. International Process and Product in Optimization Using Designed Experiments (2nd ed.). Journal of Food Science and Technology, 49(3), 726–732. Wiley, New York. https://doi.org/10.1201/9780203740774 https://doi.org/10.1111/ijfs.12357 NAJAFI, A.H., YUSOF, Y.A., RAHMAN, R.A., GANJLOO, A., LING, C.N. VIEIRA, G.S., PEREIRA, L.M., HUBINGER, M.D. 2012. Optimization of 2014. Effect of osmotic dehydration process using sucrose solution at mild osmotic dehydration process of guavas by response surface methodology and temperature on mass transfer and quality attributes of red pitaya (Hylocereus desirability function. International Journal of Food Science and Technology. polyrhizusis). International Food Research Journal, 21(2), 625–630. 47(1), 132-140.https://doi.org/10.1111/j.1365-2621.2011.02818.x NIJHUIS, H. H., TORRINGA, H. M., MURESAN, S., YUKSEL, D., LEGUIJT, WENG W., LIU, W., LIN, W. 2001. Studies on the optimum models of the dairy C., KLOEK, W. 1998. Approaches to improving the quality of dried fruit and productKou Woan Lao using response surface methodology. Asian Australasian vegetables. Trends in Food Science & Technology, 9(1), 13–20. Journal of Animal Sciences, 14(10),1470- https://doi.org/10.1016/S0924-2244(97)00007-1 1476. http://dx.doi.org/10.5713/ajas.2001.1470 ORSAT, V., CHANGRUE, V., RAGHAVAN, G.S.V.2006. Microwave drying of YADAV, B. S., YADAV, R. B., JATAIN, M. 2012. Optimization of osmotic fruits and vegetables. Stewart Postharvest Review, 2(6), 4– dehydration conditions of peach. Journal of Food Science and Technology, 49(5), 9. http://dx.doi.org/10.2212/spr.2006.6.4 547–555. http://dx.doi.org/10.1007/s13197-011-0298-z RASTOGI, N. K., RAGHAVARAO, K. S. M. S. 1997. Water and solute ZAPATA, J.E.,CIRO,G.L., MARULANDA, P. 2016. Optimization of pulsed diffusion coefficients of carrot as a function of temperature and concentration vacuum osmotic dehydration of the cape gooseberry (Physalis peruviana L.) during osmotic dehydration. Journal of Food Engineering, 34(4), 429–440. using response surface methodology. Agronomia Colombiana, 34(2), 228-238. https://doi.org/10.1016/S0260-8774(98)80034-4 http://dx.doi.org/10.15446/agron.colomb.v34n2.54920 Gandolfi, O. R. R., Goncalves, G. R. F., Bonomo, R.C.F., Fontan, R. da C. I. 2018. Sorption equilibrium and kinetics of thin-layer drying of green bell peppers. Emirates Journal of Food and Agriculture, 30(2):137-143. https://doi.org/10.9755/ejfa.2018.v30.i2.1606 SHISHIR, M. R. I., TAIP, F. S., AZIZ, N. A., TALIB, R. A., SARKER, M. S. H. 2016. Optimization of spray drying parameters for pink guava powder using RSM. Food Science and Biotechnology Journal, 25(2): 461–468. https://doi.org/10.1007/s10068-016-0064-0 SAGAR, V. R., KUMAR, S. P. 2010. Recent advances in drying and dehydration of fruits and vegetables: A review. http://dx.doi.org/10.1007/s13197-010-0010-8 SCHWEIGGERT, R.M.,MEZGER,D., SCHIMPF,F.,STEINGASS, C.B., CARLE, R. 2012. Influence of chromoplast morphology on carotenoid bioaccessibility of carrot, mango, papaya, and tomato. Food Chemistry, 135(4),2736-2742. https://doi.org/10.1016/j.foodchem.2012.07.035 SHETE, Y. V., CHAVAN, S. M., CHAMPAWAT, P. S., JAIN S. K. (2018). Reviews on osmotic dehydration of fruits and vegetables. Journal of Pharmacognosy and Phytochemistry, 7(2), 1964–1969. SHI, J., LE MAGUER, M. 2002. Osmotic dehydration of foods: mass transfer and modeling aspects. Food Reviews International, 18(4), 305–335. https://doi.org/10.1081/FRI-120016208 REZAUL, M. S. I., KARIM, N., BAO, T., GOWD, V., DING, T., SUN, C., CHEN, W. (2019). Cold plasma pretreatment-A novel approach to improve the hot air drying characteristics, kinetic parameters and nutritional attributes of shiitake mushroom. Drying Technology, 1–17. https://doi.org/10.1080/07373937.2019.1683860 SINGH, B., KUMAR, A., GUPTA, A. K. 2007. Study of mass transfer kinetics and effective diffusivity during osmotic dehydration of carrot cubes. Journal of Food Engineering, 79(2), 471–480. https://doi.org/10.1016/j.jfoodeng.2006.01.074 SINGH, B., PANESAR, P. S.,NANDA, V. 2008. Optimization of osmotic dehydration process of carrot cubes in sucrose solution. Journal of Food Process Engineering, 31(1), 1–20. https://doi.org/10.1016/j.foodchem.2010.04.075 SINGH, B., PANESAR, P. S., NANDA, V., KENNEDY, J. F. 2010. Optimisation of osmotic dehydration process of carrot cubes in mixtures of sucrose and sodium chloride solutions. Food Chemistry, 123(3), 590–600. https://doi.org/10.1016/j.foodchem.2010.04.075 Singh, R., Giri, S. K., Kotwaliwale, N. 2014. Shelf-life enhancement of green bell pepper (Capsicum annuum L.) under active modified atmosphere storage. Food Packaging and Shelf Life, 1(2), 101–112. https://doi.org/10.1016/j.fpsl.2014.03.001 Stat-Ease, I. (2007). Design Expert Software, Educational Version 7.0.3. Wiley Publishing. SUTAR, P. P., PRASAD, S. 2011. Optimization of osmotic dehydration of carrots under atmospheric and pulsed microwave vacuum conditions. Drying Technology, 29(3), 371–380. https://doi.org/10.1080/07373937.2010.497955 TALENS, P., MARTÍNEZ-NAVARRETE, N., FITO, P., CHIRALT, A. 2002. Changes in optical and mechanical properties during osmodehydrofreezing of kiwi fruit. Innovative Food Science & Emerging Technologies, 3(2), 191– 199.https://doi.org/10.1016/S1466-8564(02)00027-9 TELIS, V. R. N., MURARI, R. C. B. D. L.,YAMASHITA, F. 2004. Diffusion coefficients during osmotic dehydration of tomatoes in ternary solutions. Journal of Food Engineering, 61(2), 253–259. https://doi.org/10.1016/S0260- 8774(03)00097-9 TORRES, J. D., TALENS, P., CAROT, J. M., CHIRALT, A., ESCRICHE, I. 2007. Volatile profile of mango (Mangifera indica L.) as affected by osmotic dehydration. Food Chemistry, 101(1), 219–228.
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GARLIC-GINGER AS POTENTIAL BIO-PRESERVATIVE IN FERMENTED MAIZE AND SORGHUM PASTES
Abiola Folakemi Olaniran1,2*, Sumbo Henrietta Abiose3, and Adeniran Adekanmi Hezekiah 3
Address(es): Abiola Folakemi Olaniran, *1Landmark University, College of Pure and Applied Sciences, Department of Microbiology, 251101, Omu-Aran, Kwara State, +2348034961968: 2Landmark University, College of Agricultural Sciences, Department of Food Science and Nutrition, 251101, Omu-Aran. 3Obafemi Awolowo University, Faculty of Technology, Department of Food Science and Technology, 220282, Ile-Ife, Osun State.
*Corresponding author: [email protected] doi: 10.15414/jmbfs.2020.10.3.467-473
ARTICLE INFO ABSTRACT
Received 24. 6. 2019 Garlic and ginger are natural spices with potentials as biopreservatives and allied health benefits. Fermented pastes either from maize or Revised 14. 8. 2020 sorghum have a shelf life of fewer than 10 days except when refrigerated. In this study, garlic and ginger were added separately and in Accepted 24. 9. 2020 combinations to the fermented pastes prepared from maize and sorghum grains with a view to extending its shelf life resulted in 7 Published 1. 12. 2020 treatments. During storage for 4 weeks at ambient and refrigeration temperatures, the microbial load was enumerated and isolates identified using conventional methods. Physicochemical properties and shelf-life of the paste were also evaluated. Prominent in the fermented pastes during storage were 8 species of lactic acid bacteria and yeast (Saccharomyces cerevisiae and Saccharomyces rouxii). Regular article Garlic inhibited the growth of Candida utilis, Candida mycoderma, Candida tropicalis, Candida krusei, and Rhodotorula glutinis in paste during storage. Bio-preserved paste using 4% garlic with 2% ginger had the best preservative effect on total viable counts (8.29- 11.30 CFU/g), lactic acid bacteria (8.3-9.70 CFU/g) and yeast (4.69-9.45 CFU/g) counts. The study established that fermented pastes produced from either maize or sorghum can be effectively bio preserved using garlic, ginger, and garlic-ginger at 2 or 4 % for 4 weeks without spoilage at ambient temperature (27± 2oC); thus, extending its shelf life.
Keywords: Fermented paste, Biopreservatives, Garlic, Ginger, Maize, Sorghum, Shelf life
INTRODUCTION maize and sorghum with garlic and ginger as biopreservative at ambient and refrigeration temperatures. Quality protein maize (QPM) varieties are being encouraged in developing countries with maize as major constituents of their diets compared to the MATERIAL AND METHODS conventional maize (Gunaratna, et al., 2019). Different varieties are used for the production of several traditional foods especially in Africa such as ogi, The main Preparation of powder garlic and ginger diets of poor people are cereals with scanty protein sources like eggs and meat (Oluwakemi and Omodele, 2015). Quality Protein Maize (QPM) has a Garlic and ginger powder was prepared according to Olaniran et al. (2015) from biological protein value of 80% compared with conventional maize (45%). fresh garlic bulbs and ginger rhizomes. Four hundred (400) gram of fresh garlic Besides, It has more lysine (30%), tryptophan (55%) and less leucine (38%) with bulbs and ginger rhizomes were cleaned, peeled and dried in hot air oven the potential in alleviating protein malnutrition (Maseta et al., 2017). The (Gallenkamp, UK) at 65oC for 12h. Dried samples were pulverized with a grinder superiority of nutritional assessment on consumption of quality protein maize by (Marlex Appliances PVT, India). The garlic and ginger powdered samples were human being above other varieties has been demonstrated (Tandzi et al., 2017). then sieved in a mesh (50µm). Sorghum is the world's fifth cereals food crop. A good source of starch and protein, its gluten-free serves as an alternative for gluten intolerance people Production of biopreserved ogi paste (Kulamarva et al., 2009). Fermentation is a traditional, age-old technique of transforming grains generally, into diverse forms of food that constitute the daily The paste produced from clean sorghum (red variety) and quality protein maize diets of most African populations. This processing technique has been well grains from the Institute of Agricultural Research and Training (I.A.R.T.), documented to improve shelf life, nutrient bioavailability and health benefits Ibadan, Nigeria. The grains soaked in clean water for 72h individually, drained, (Olaniran and Abiose, 2019). Some of the locally fermented products are fura, wetly milled to smooth paste in attrition mill using Thomas Wiley Model ED-5 ogi and tings. Most of these fermented products especially ogi has relatively short (Akanbi et al., 2003, Olaniran and Abiose, 2018). The smooth paste divided to shelf life of 10 days at room temperature depending on the moisture content and 8 portions (800g each); Fermented paste without biopreservative served as microorganisms present (Olaniran et al., 2019). Refrigeration is a viable option control and labeled PWB. The garlic and ginger powder added to the slurry in when the power supply is regular for preservation if the family can afford the bill. different concentrations (2 and 4%). Other portions labeled as follows: BPG1 (2% Therefore there is a need to extend their shelf life to enhance its availability on garlic), BPG2 (4% garlic); BPG3 (2% ginger), BPG4 (4% ginger), BPG5 (2% the shelf and reduce the rigor of frequent production in small batches. garlic with 2% ginger), BPG6 (2% garlic with 4% ginger), and BPG7 (4% garlic Biopreservation is important in the food industry. Ginger powder compared well with 2% ginger). The mixtures were steadily homogenized in containers with a with potassium sorbate, citric acid, and sodium metabisulphite (Singh, 2018). glass rod and fermented at ambient temperature (27± 2oC) for 24h. It was Also, Olaniran et al. (2015) reported that garlic possesses antimicrobial decanted and packaged in plastic containers stored at ambient temperature (27 ± activities in food. Garlic and ginger satisfy the questions of safety and generally 2 °C) and refrigeration (4 ± 1oC) temperatures. Samples were selected at random regarded as safe (GRAS) (Olaniran et al., 2020). They are indispensable in the in 4 weeks in replicates for futher studies preparation of daily food and reported to possess compounds with various benefits (Tattari et al., 2013). Garlic and ginger are gaining increasing interest because of relatively safe status, acceptance by consumers, and a potential alternative to chemical preservatives (Nath et al., 2014). Hence, the study was designed in view of extending the shelf life of fermented pastes produced from
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Microbial Analyses of 3.0% Ninhydrin solution in 2-methoxyethanol were added. The tubes were heated in a water bath for 15 min, 10ml of isopropyl alcohol: water mixture (1:1) Enumeration of Microbes was rapidly added then cooled to room temperature. The Optical Density (O.D) of the solution was read at 570 nm in a spectrophotometer (Model SP9, Enumeration of microbes by serial dilution of fermented paste was carried out PyeUnican UK). The concentration of free amino acids in the extract was during storage. Mixing of the paste (5g) with 45ml of maximum recovery extrapolated from a standard curve of known concentrations of glycine. diluents (MRD) resulting in 10-1 dilution. The stock was further diluted till a preferred dilution was obtained. From each dilution, 1ml was pipetted into sterile Determination of total reducing sugars petri dish prior to adding 20mL each of molten Nutrient agar (NA) for enumeration microorganisms. Molten agar of de Man Rogosa-Sharpe (MRS) The total reducing sugar content was determined using the dinitrosalicyclic acid was added for Lactic acid bacteria (LAB) count and Potato dextrose agar (PDA) (DNSA) reagent method described by Adepoju et al. (2016). One (1) milliliter of for yeast and mold count (to inhibit bacterial growth, 0.5ml of 3 mg/ml ethanolic extract was dispensed into test tubes and 2 ml of DNSA was added. The Streptomycin was incorporated). Petri dish in triplicates were incubated at 37oC mixture boiled for 5 min, rapidly cooled under running water and 7ml of distilled for 24h for the total viable count (TVC) and LAB count while for yeast and mold water was added. The absorbance was read at 540 nm in a UV Spectrophotometer count media were incubated at 28 ± 2oC for 72h. The microbial load in paste as (Model SP9, PyeUnican UK) against reagent blank. Results were extrapolated storage progressed was determined to compute distinct colonies using a colony from concentrations of a standard glucose calibration curve. counter. Incubated Petri dish with 25-250 colonies was counted and numbers of colonies multiplied with reciprocal dilution factor (Harrigan, 1998; APHA, 2015).
Characterization and Identification of Isolates Effect of Biopreservative on individual sugars in fermented paste
Distinct colonies were streaked on solidified agar repeatedly to obtain pure Thin layer chromatography described by Adeniran and Abiose (2012) in the isolate and subjected to microscopic examination for identification. The isolates identification of individual sugars was used. Newly prepared standard sugars were identified based on cultural, morphological characteristics and biochemical containing 0.5 % w/v maltose, glucose, and galactose were stored in bottles. assays according to Taubeneck (2007), Wood, and Holzapfel (1995). Shape and Micropipette was used to spot the standard sugar solution on chromatogram sheet elevation of the colonies were examined, Gram reaction, cell morphology and (pre-coated silica gel adsorbent for thin-layer chromatography) and the extracts. cell arrangement of the isolates were observed under a microscope. Identification Prepared solvent system mixing 60 % ethyl acetate, 15 % glacier acetic acid, 15 of yeast and bacteria was done using conventional method carrying out catalase % ethanol and 10 % distilled water together. The dried spotted plate was test, oxidase test, Gram staining, sugar fermentation and production of carbon immersed in the saturated chromatography tank containing the solvent system, dioxide. Relevant scheme were consulted (Harrigan, 1998; Stöckheim, 2007; uprightly positioned and allowed to develop for 1 h. The developed sheet was Taubeneck, 2007; Wood and Holzapfel, 1995) for the identification of isolates gently removed and dried at room temperature. The detection reagent prepared by strain. mixing naphthol resorcinol (20 mg), 90% ethanol (10 ml), and concentrated sulphuric acid (0.2 ml). Detection reagent was sprayed on the dried plate, dried in Determination of Titratable acidity and pH the oven at 100oC for 10 min resulting in defined and distinct spots on dried chromatogram sheet. The total titratable acidity (TTA) of fermented paste was done using 25 ml of the slurry and 3 drops of phenolphthalein as indicator and titrated against 0.1N RESULTS AND DISCUSSION Sodium hydroxide (NaOH) to achieve pink color. The acidity of paste was calculated as lactic acid with conversion factor 0.09. pH meter (Corning Scholar Effects of addition of biopreservative on fermented paste 425, UL Laboratories, Shenzhen, China) was used for determination of pH values. Buffer 4.0 and 7.0 used to calibrate the pH meter and the electrode The total viable counts (TVC) of PWB (paste without preservative) increased probes sanitized by swabbing with 90% ethanol before dipping in paste (AOAC, ranging from 11.98-16.97 CFU/g (maize) and 11.99-15.99 CFU/g (sorghum) 2010). during storage (Table 1). The viable counts of BPG1 (bipreserved paste with 2% garlic) and BPG2 (4% garlic) gradually increased from 8.45-9.81 CFU/g and 8.35 Preparation of extracts for total free amino acid, total reducing sugars, total to 11.59 CFU/g respectively in 4 weeks at ambient shelf life. The viable counts soluble sugars and individual sugars assay recorded in all biopreserved pastes in week 4 were lower than counts in week 2 compared to that of PWP (control) at week 2. The preservative effect in BPG1 in Extraction of the paste was carried out according to Omafuvbe (2006). Paste week 4 (11.68 CFU/g) was comparable to that of BPG3 in week 2 (11.71CFU/g) (5g) was weighed into 250ml conical flask and 50ml of 80% ethanol (v/v) was in pastes produced from maize at ambient temperature as documented in table 1. added. The suspension was mixed and 10ml of petroleum ether was added to Both BPG2 (11.59 CFU/g) and BPG5 (11.40 CFU/g) showed the nearly equivalent extract the fat in paste. The ethanol- petroleum ether was stirred using magnetic preservative effect in week 4. The use of garlic- ginger in combination with stirrer at room temperature for 30 min and centrifuged at 5000 rpm for 30 refrigeration further reduced total viable counts of preserved paste during the 4 minutes. The petroleum ether phase was discarded and clear ethanolic phase was weeks of storage by extending the shelf life of the paste by 200%. Also, TVC of collected and used for further analysis. BPG7 was stable at refrigeration temperature during storage. The lowest TVC count (1.27 CFU/ g) was recorded during storage in BPG7 (preserved paste with Determination of total free amino acid 4% garlic-2% ginger) which was prepared from maize (supplementary 1).
Ninhydrin colorimetric method according to Omafuvbe (2006) to 1ml of the extract in labeled test tube, 0.5 ml of cyanide acetate buffer (pH 5.4) and 0.5ml
Table 1 Total Viable Count (CFU/ g) of biopreserved fermented pastes at ambient temperature Time PWB BPG BPG BPG BPG BPG BPG BPG (weeks) 1 2 3 4 5 6 7 Maize 0 11.98±0.11a 8.45±0.10c 8.35±0.13c 8.65±0.14c 8.62±0.10c 8.90±0.10b 8.91±0.10b 8.29±0.10d 2 15.56±0.10a 9.81±0.11c 9.77±0.15c 11.92±0.12b 11.85±0.13b 9.69±0.03d 9.40±0.15e 9.32±0.11f 4 16.97±0.10a 11.68±0.15b 11.59±0.13d 11.89±0.10c 11.71±0.15b 11.40±0.15b 11.34±0.12b 11.30±0.15b Sorghum 0 11.99±0.15a 11.63±0.10c 11.57±0.10c 11.76±0.12b 11.65±0.10c 11.43±0.10d 11.34±0.15c 11.45±0.11d 2 13.94±0.12a 11.91±0.15b 11.87±0.11c 11.96±0.15b 11.94±0.12b 11.64±0.12d 11.49±0.12c 11.54±0.15d 4 15.99±0.11a 13.40±0.15c 11.94±0.15d 13.81±0.12b 13.67±0.10c 11.93±0.10d 11.78±0.11c 11.86±0.15de Means followed by different superscripts are significantly different across rows (p < 0.05). Keys: PWB (paste without biopreservative), BPG1 (2% garlic), BPG2 (4% garlic); BPG3 (2% ginger), BPG4 (4% ginger), BPG5 (2% garlic-2% ginger), BPG6 (2% garlic-4% ginger), and BPG7 (4% garlic-2% ginger).
Lactic acid bacteria counts for all samples during storage for 4 weeks ranged (13.77 CFU/g) and BPG4 (13.80 CFU/g) observed in fermented paste produced from 8.30 to 16.96 CFU/g (maize) and 8.40-13.93 CFU/g (sorghum) at ambient using sorghum; at ambient temperature showed no significant difference at the temperature. LAB counts of biopreserved paste increased all through the 4 end of week 4. Also, the use of only garlic and garlic-ginger showed stabilizing weeks of storage (Table 2). The LAB counts of PWP (13.82 CFU/g), BPG3 effect on lactic acid bacteria growth in 4 weeks. The significant distinction in
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LAB counts of garlic-ginger as biopreservatives as the storage of paste yeast and mould count during storage. Ajoene, a garlic-derived sulfur-containing progressed was noticeable. This demonstrated more antimicrobial activity of compound reported by Olaniran et al. (2020) to be responsible for this garlic against gram-positive bacteria and gram-negative bacteria. observation. Low LAB and yeast count in preserved pastes at both temperatures Yeast and mould counts in PWP ranged from 8.81-13.76 CFU/g (maize) and may be due to the antimicrobial properties of garlic and ginger (Asimi et al., 8.39-15.68 CFU/g (sorghum). Addition of garlic and garlic-ginger as 2017). Similar reports by Oladipo and Jadesimi, (2013) and Olaniran et al. biopreservatives significantly reduced yeast and mould counts in 4 weeks at (2015) during the storage of food products preserved singly with either ginger or ambient temperature from 13.76 and 15.68 CFU/g to range of 9.45-11.59 CFU/g garlic was documented. BPG6 (4% garlic-2% ginger) had the best effect on the and 11.39-13.92 CFU/g in pastes produced form maize and sorghum total viable, Lactic acid bacteria and yeast count during storage of paste produced respectively. Yeast and mould counts in BPG6 were relatively constant (4.86-4.95 from maize and sorghum. and 5.91-5.99 CFU/g) in paste from maize and sorghum at refrigeration temperature (Table 3). Garlic was found to be more effective than ginger against
Table 2 Lactic Acid Bacteria Count (CFU/ g) of biopreserved fermented pastes at ambient temperature Time PWB BPG BPG BPG BPG BPG BPG BPG (weeks) 1 2 3 4 5 6 7 Maize 0 9.34±0.15bc 8.76±0.10b 9.43±0.01b 9.13±0.11c 9.63±0.15a 8.90±0.10d 8.46±0.03e 8.30±0.02f 2 13.92±0.10a 9.70±0.15d 9.17±0.12f 11.60±0.05b 9.99±0.01c 9.34±0.02e 9.03±0.05g 9.01±0.03g 4 16.96±0.02a 9.94±0.04d 9.80±0.03c 11.95±0.01b 11.45±0.03c 9.74±0.01e 9.62±0.06f 9.70±0.01e Sorghum 0 11.60±0.11a 9.43±0.12ab 8.51±0.15d 9.91±0.10b 9.82±0.10b 8.64±0.11c 8.40±0.15e 8.48±0.15f 2 13.93±0.10a 11.68±0.13c 11.63±0.01c 11.80±0.03b 11.89±0.05b 10.73±0.05e 10.45±0.01e 10.69±0.05d 4 13.82±0.05a 11.86±0.03b 11.76±0.05c 13.77±0.05a 13.80±0.01a 11.71±0.08c 11.58±0.02d 11.67±0.00d Means followed by different superscripts are significantly different across rows (p < 0.05). Keys: PWB (paste without biopreservative), BPG1 (2% garlic), BPG2 (4% garlic); BPG3 (2% ginger), BPG4 (4% ginger), BPG5 (2% garlic-2% ginger), BPG6 (2% garlic-4% ginger), and BPG7 (4% garlic-2% ginger).
Table 3 Yeast count (CFU/ g) of biopreserved fermented pastes at ambient temperature Time PWB BPG1 BPG2 BPG3 BPG4 BPG5 BPG6 BPG7 (weeks) Maize 0 8.81±0.03a 5.36±0.01d 5.27±0.00c 6.63±0.02b 5.78±0.01b 4.74±0.04d 4.86±0.03d 4.69±0.00d 2 11.60±0.03a 8.74±0.03c 8.56±0.03c 9.86±0.03b 9.60±0.03b 8.45±0.03d 8.40±0.03d 8.30±0.03e 4 13.76±0.03a 9.86±0.03c 9.81±0.03c 11.53±0.03b 11.39±0.03b 9.72±0.03e 9.57±0.03d 9.45±0.03d Sorghum 0 8.39±0.01a 6.67±0.02b 6.48±0.01b 6.61±0.05b 6.73±0.03b 6.32±0.00c 5.91±0.08c 5.98±0.05c 2 11.87±0.03a 8.85±0.03c 8.81±0.03c 9.94±0.03b 9.91±0.03b 8.83±0.03c 8.36±0.03d 8.74±0.03c 4 15.68±0.03a 11.93±0.03c 11.73±0.03c 13.92±0.03b 13.85±0.03b 11.67±0.03c 11.39±0.03d 11.50±0.03d Means followed by different superscripts are significantly different across rows (p < 0.05). Keys: PWB (paste without bio-preservative), BPG1 (2% garlic), BPG2 (4% garlic); BPG3 (2% ginger), BPG4 (4% ginger), BPG5 (2% garlic-2% ginger), BPG6 (2% garlic-4% ginger), and BPG7 (4% garlic-2% ginger).
Identification and occurrence pattern of microorganisms in fermented paste Candida krusei, and Rhodotorula glutinis were inhibited during storage in pastes having garlic. Garlic has been reported to have high anti-candida activity and Eight lactic acid bacteria isolates were identified as Lactobacillus plantarum, fungicidal effects against Rhodotorula (Ali and Mustafa, 2009; Ogunsakin et Lactobacillus amylovorus, Leuconostoc mensenteroides, Lactobacillus al., 2015). delbrueckii, Lactobacillus brevis, Lactobacillus fermentum, Pediococcus pentosaceus, and Pediococcus acidilactici. Lactobacillus plantarum was Total Titratable Acidity and pH of paste dominant throughout the period of storage at ambient and refrigeration temperature in paste produced from sorghum. Lactobacillus amylovorus was The total titratable acidity (TTA) value of the pastes increased at ambient and present for first 2 weeks at ambient temperature and throughout at refrigeration refrigeration temperatures during the 4 weeks of storage respectively. Control temperature. Leuconostoc mensenteroides was also present throughout storage at sample from paste produced from sorghum had the highest titratable acidity refrigeration temperature but only in the first week at ambient temperature. (0.25) while lowest was observed in BPG7 (0.15) in week 4 at ambient Lactobacillus brevis, Pediococcus pentosaceus, and Pediococcus acidilactic temperature. The TTA and pH of BPG6 and BPG7 were stable during storage isolated in week 2 at both temperatures as presented in Table 4. They have been (figures 1 and 2). BPG7 total titratable acidity and pH were stable at refrigeration reported as major lactic acid bacteria isolated from fermented cereal dough temperature. The variations in acid tolerance of microorganisms could be (Adesulu-Dahunsi et al., 2017; Bello et al., 2018; Okoronkwo, 2014; attributed to their relative ATPase activities as the pH decreases (Wakil and Olayiwola et al., 2017; Wakil and Ajayi, 2013). Yeast isolates were identified Ajayi, 2013). Effective inhibition of microorganisms depends on numbers of as Saccharomyces cerevisiae, Saccharomyces rouxii, Candida utilis, Candida lactic acid bacteria sufficient to decrease the pH rapidly to levels that inhibit mycoderma, Candida tropicalis, Candida krusei, Hansenula anomala, and growth of pathogen. Decreased in pH and increase in TTA in the study was in Rhodotorula glutinis (Table 5). Saccharomyces cerevisiae and Saccharomyces agreement with Oyedeji et al. (2013). rouxii were dominant for 4 weeks as shown in the occurrence pattern of yeast isolates. The growth of Candida utilis, Candida mycoderma, Candida tropicalis,
Table 4 Morphological and Biochemical Characteristics of Bacteria Isolates from of biopreserved fermented pastes Test Isolates 1 2 3 4 5 6 7 8 9 10 Morphology Cocc Rod Rod Rod Cocci Rod Rod Cocci Cocci Rod i Colour of growth Yell Cream Cream Cream Cream Cream Cream Cream Cream Cream ow Gram reaction + + + + + + + + + + Catalase test + + ------Growth at: 15 oC - + + - + - + + + - 45 oC + + - + - + - - + + Production of CO2 + Nd - - + - + - - - Starch + - - + ------Nitrate ------+ Nd Nd + Dextran production + + - - + - - - - -
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NH3 from arginine - + - - Nd - + + - + fermentation of: Glucose + + - - + + + + + + Lactose + - + - + + + + + + Maltose + + + + + + + + Arabinose - - + + + - + - + + Trehalose + - + + + + - + + + Salicin - - + + + - - + + - Sucrose + + + - + + + - - + Raffinose - - + - + - + + + + Mannitol + + Nd - + - - + + Nd Xylose - + + - + - + - + + Fructose + + Nd - + Nd + + Galactose + + + + + - + + + + Citrate - + + Probable identity of Corynebacterium M. L. L. L. P. P. L. L. plantarum L. brevis the organism spp. luteus amylovorus mensenteroides delbrueckii pentosaceus acidilactici fermentum
Table 5 Morphological and Biochemical Characteristics of Yeast Isolates from of biopreserved fermented pastes Test Isolates 1 2 3 4 5 6 7 8 Morphology: Colour Cream Cream Red Cream Cream Yellow Cream Grey Shape Ovoid Ovoid Ovoid Spherical Ovoid Cylindrical Cylindrical Cylindrical Reproduction Polar Polar Multilateral Polar Multilateral Multilateral Multilateral Multilateral (Budding) Budding Budding Budding Fermentation:
Glucose + + - + + + + - Sucrose + - - + + - + - Maltose + +(w) - +(w) + - - - Galactose + - - + + - - - Raffinose + - - +(w) - - + - Lactose ------Sugar assimilation Glucose + + + + + + + + Sucrose + - + + + - + - Maltose + + + + + - + - Galactose + + + + + - - - Raffinose + - + + - - + - Lactose ------Trehalose - - + + - - - - Pellicle - - - + + + - + Nitrate assimilation - - - + - - + - Probable identity of Saccharomyces Saccharomyces Rhodotorula Hansenula Candida Candida krusei Candida utilis Candida mycoderma organism Cerevisiae Rouxii glutinis anomala tropicalis Key: -: negative; +: positive; w: weak
0,45 (b) Sorghum 0,45 (a) Maize 0,4 0,4
0,35
0,35 0,3 0,3 0,25 0,25
0,2 0,2 PWB BPG1 BPG2
Titratable Acidity (%) Acidity Titratable 0,15
0,15 Titratable Acidity (%) Acidity Titratable BPG3 BPG4 BPG5 PWB BPG1 BPG2 0,1
Total 0,1
BPG3 BPG4 BPG5 Total 0,05 0,05 BPG6 BPG7 BPG6 BPG7 0 0 0 2 4 0 2 4 Storage Period (weeks) Storage Period (weeks)
Figure 1 Total titratable acidity of Biopreserved fermented pastes at Ambient Temperature (a) maize, (b) Sorghum
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120 3,4
(a) Maize PWB (b) Sorghum
3,2 BPG1 100
3 BPG2 80
2,8 BPG3
pH 60 2,6 BPG4
2,4 BPG5 40 PWB BPG1 BPG2 BPG6 2,2 20 BPG3 BPG4 BPG5 BPG7 2 Total Free Amino Acid (mg glycine/ml) (mg Acid Amino Free Total BPG6 BPG7 0 2 4 0 Storage Period (weeks) 0 2 4
Figure 3 Total free amino acid of biopreserved fermented pastes at Ambient 4 (b) Sorghum Temperature 3,8 Total Reducing Sugar Content of fermented paste 3,6 3,4 The total reducing sugar (TRS) of paste during storage ranged from 6.21 - 41.87 mg/ml (maize) and 19.97-51.32 mg/ml (sorghum) at ambient temperature. 3,2 Relatively stable total reducing sugar content was documented in BPG4 and BPG5 (Figures 4). PWB (Control paste from maize) had the lowest total reducing 3 sugar (6.21 mg/ml) in 4 weeks while BPG from sorghum had the highest (51.32 PWB BPG1 BPG2 5 mg/ml) at ambient temperature. Increased total soluble sugar may be due to
pH 2,8 maximum breaking down of starch substrates to simpler sugars by amylolytic 2,6 BPG3 BPG4 BPG5 enzymes produced during fermentation (Adesulu et al. 2014; Okafor et al. 2018; Oyarekua and Adeyeye, 2009). Low reducing sugar content in fermented 2,4 pastes has been reported as a good attribute because methods of preparing 2,2 BPG6 BPG7 weaning foods involve heat processing by Oyerekau and Adeyeye (2009).
2 45 0 2 4
40 Storage Period (weeks) (a) Maize
Figure 2 pH of Biopreserved fermented pastes at Ambient Temperature (a) 35 maize, (b) Sorghum 30
The total free amino acid of fermented paste 25
The total free amino acid (TFAA) content of pastes ranged from 44.96 to 73.70 20 mg/ml and 37.18-102.56 mg/ml for pastes form maize and sorghum at ambient temperature respectively. Stable TFAA observed during storage of BPG1, BPG2, 15 PWB BPG1 BPG2 BPG , BPG and BPG at ambient temperature (Figure 3). The TFAA content of 4 5 7 10 PWB increased throughout the 4 weeks of storage at a refrigeration temperature BPG3 BPG4 BPG5 from 44.69 to 50.62 mg/ml and 37.18 to 51.00 mg/ml in pastes form maize and 5 BPG6 BPG7 sorghum respectively. Rojas-Molina et al. (2008) reported a similar observation
of increased in free amino acid. 0 Total Reducing Sugar (mg glucose/ml) (mg Sugar Reducing Total 0 2 4 Storage period (Weeks) 80 PWB (a) Maize 70 BPG1 60 BPG2 60 50 BPG3 (b) Sorghum 50 40 40 BPG4 BPG5 30 30 BPG6 20 BPG7 20 PWB BPG1 BPG2 10 BPG3 BPG4 BPG5
10
Total Free Amino Acid (mg glycine/ml) (mg Acid Amino Free Total 0 BPG6 BPG7 0 2 4 0 Storage period (Weeks) glucose/ml) (mg Sugar Reducing Total 0 2 4 Storage period (Weeks)
Figure 4 Total reducing sugar of biopreserved fermented pastes at Ambient Temperature
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12, 55–69. https://doi.org/10.1080/10942910802252148 Identification of individual sugars in fermented paste extract Maseta, E., Mosha, T. C., Nyaruhucha, C., Laswai, H. 2017. Nutritional quality of quality protein maize-based supplementary foods. Nutrition and Food Science, Individual sugars were not present in all paste extracts at week 0. However in 47(1), 42–52. https://doi.org/10.1108/NFS-04-2016-0042 paste produced from maize; glucose was the main sugar present in PWB, BPG1, BPG2, and BPG4, BPG5, BPG6 and BPG7 at week 4 at ambient temperature. Also Nath, S., Chowdhury, S., Dora, K., Sarkar, S. 2014. Role of Biopreservation in in pastes made from sorghum, sucrose and glucose with calculated refractive Improving Food Safety and Storage. Journal of Engineering Research and index (Rf value) 0.31 and 0.41 were present in BPG6 in 4 weeks at ambient Applications www.Ijera.Com, 4(1), 26–32. www.ijera.com temperature. Fructose with calculated refractive index (Rf value) of 0.46 was Ogunsakin, O. A., Banwo, K., Ogunremi, O. R., Sanni, A. I. 2015. identified in BPG3, BPG5, BPG6, and BPG7 in 4 weeks at ambient temperature. Microbiological and physicochemical properties of sourdough bread from Individual sugars were not present in all paste extracts stored at refrigeration sorghum flour. International Food Research Journal. temperature. Glucose reported as one of the major monosaccharide present in Okafor, U. I., Omemu, A. M., Obadina, A. O., Bankole, M. O., Adeyeye, S. A. significant quantity in cereal grains such as maize and sorghum by Omemu O. 2018. Nutritional composition and antinutritional properties of maize ogi (2018) and Akinrinola et al. (2014). cofermented with pigeon pea. Food Science and Nutrition, 6(2), 424–439. https://doi.org/10.1002/fsn3.571 CONCLUSION Okoronkwo, C. U. 2014. Isolation and Characterization of Lactic Acid Bacteria involved in the Fermentation of Millet and Sorghum sold in Nkwo-Achara This study established that fermented paste produced from maize and sorghum market, Abia State.IOSR Journal of Environmental Science, Toxicology and can be effectively biopreserved with garlic-ginger for 4 weeks at ambient Food Technology, 8(9), 42-45. temperature without deteriorating. The most effective combination was 4% garlic www.iosrjournals.orgwww.iosrjournals.org42%7C and 2% ginger in maize and sorghum as biopreservatives. Thus, garlic, ginger Oladipo, I. C., Jadesimi, P. D. 2013. Microbiological Analysis and Nutritional addition are highly recommended as biopreservatives in fermented paste for Evaluation of West African soft cheese (wara) produced with different extending shelf life in our bid to sustain its availability in remote communities preservatives. American Journal of Food and Nutrition, 3(1), 13–21. with inconsistent power supply and probable application for industrial https://doi.org/10.5251/ajfn.2013.3.1.13.21 production. Olaniran, A. F., Abiose, S. H. 2018. Proximate and antioxidant activities of bio- preserved ogi flour with garlic and ginger. F1000Research, 7 (1936),1-13. Acknowledgments: Institute of Agricultural Research and Training (I.A.R.T.), https://doi.org/10.12688/f1000research.17059.1 Ibadan, Nigeria is appreciated for providing quality protein maize used for this Olaniran, A. F., Abiose, S. H. 2019. Nutritional evaluation of enhanced unsieved research. Ogi Paste with Garlic and Ginger. Preventive Nutrition and Food Science, 24(3), 348-356. https://doi.org/10.3746/pnf.2019.24.3.348 REFERENCES Olaniran, A. F., Abiose, S. H., Adeniran, A. H. 2015. Biopreservative Effect of Ginger (Zingiber officinale) and Garlic Powder (Allium sativum) on Tomato Akinrinola, A. Adejimi M., Idabor, V., Oliseh, C., Sanni, R., Oshagbemi, H., Paste. Journal of Food Safety, 35(4), 440–452. https://doi.org/10.1111/jfs.12193 Oyefuga, O. 2014. Dietary Fortification of Ogi (Maize slurry ) with Okra seed Olaniran, A. F., Abiose, S. H., Gbadamosi, S. O. 2019. Nutritional Quality and flour and its Nutritional value. Scholarly Journal of Agricultural Science, 4(4), Acceptability Evaluation of Ogi Flour Biofortified with Garlic and Ginger. 213–217. https://www.researchgate.net/publication/291335302 Journal of Health Science, 7, 101–109. https://doi.org/10.17265/2328- Adepoju, A. A., Abiose, S. H., Adeniran, H. A. 2016. Effect of pasteurization and 7136/2019.02.005 selected chemical preservatives on Fura de nunu during storage. African Journal Olaniran, A. F., Abu, H. E., Afolabi R. O. Okolie, C., Owolabi A., Akpor O. of Food Science and Technology, 7(8), 178–185. 2020. Comparative assessment of storage stability of Ginger-Garlic and Chemical https://doi.org/10.14303/ajfst.2016.102 preservation on Fruit juice blends. Potravinarstvo Slovak Journal of Food Adeniran, H. A., Abiose, S. H. 2012. Partial purification, characterization and Sciences 14, 88-94. https://doi.org/10.5219/1262 hydrolytic activities of amylases from Bacillus licheniformis and Aspergillus Olayiwola, J. O., Inyang, V., Bello, M. A. 2017. Bacteriological and Proximate niger cultured on agricultural residues. African Journal of Biotechnology, 11(6), Evaluation of Ginger-Fortified Fermented Maize (OGI). American Journal of 1465–1477. https://doi.org/10.5897/AJB10.2233 Food Technology, 12(6), 374–378. https://doi.org/10.3923/ajft.2017.374.378 Adesulu-Dahunsi, A. T., Sanni, A. I., Jeyaram, K. 2017. Rapid differentiation Oluwakemi, A., Omodele, I. 2015. The current status of cereals (maize, rice and among Lactobacillus, Pediococcus and Weissella species from some Nigerian sorghum) crops cultivation in Africa: Need for integration of advances in indigenous fermented foods. https://doi.org/10.1016/j.lwt.2016.11.007 transgenic for sustainable crop production. International Journal of Agricultural Adesulu, A., T., Awojobi, K., O. 2014. Enhancing sustainable development Policy and Research, https://doi.org/10.3923/ajft.2017.374.378 through indigenous fermented food products in Nigeria. African Journal of Omafuvbe, O. B. 2006. Effect of salt on the fermentation of soybean (Glycine Microbiology Research, 8(12), 1338–1343. max) into daddawa using Bacillus subtilis as starter culture. African Journal of https://doi.org/10.5897/AJMR2013.5439 Biotechnology , 5(10), 1001–1005. https://doi.org/10.5897/ajb06.169 Akanbi, C. T., Ade‐Omowaye, B. I., Ojo, A., Adeyemi, I. A. 2003. Effect of Omemu, A. M. 2011. Fermentation dynamics during production of ogi, a Processing Factors on Rheological Properties of Ogi. International Journal of Nigerian fermented cereal porridge. Report and Opinion, 3(4), 8–17. Food Properties, 6(3), 405–418. https://doi.org/10.1081/JFP-120020118 http://www.sciencepub.net/report Ali, A. A., Mustafa, M. M. 2009. Isolation, Characterization and Identification Omemu, A. M., Okafor, U. I., Obadina, A. O., Bankole, M. O., Adeyeye, S. A. of Lactic Acid Bacteria from Fermented Sorghum Dough used in Sudanese Kisra O. 2018. Microbiological assessment of maize ogi cofermented with pigeon pea. Preparation. Pakistan Journal of Nutrition, 8(11), 1814–1818. Food Science and Nutrition, 6(5), 1238– 1253. https://doi.org/10.1002/fsn3.651 http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.604.1248&rep=rep1&t Oyarekua, M. A., Adeyeye, E. I. 2009. Comparative Evaluation of the Nutritional ype=pdf Quality, Functional Properties and Amino Acid Profile of Co-Fermented AOAC. 2010. Official methods of analysis of AOAC International. Gaithersburg Maize/Cowpea and Sorghum/Cowpea Ogi as Infant Complementary Food. Asian MD.: AOAC International. https://www.worldcat.org/title/official-methods-of- Journal of Clinical Nutrition, 1(1), 31–39. analysis-of-aoac-international/oclc/649275444 https://doi.org/10.3923/ajcn.2009.31.39 APHA 2015. Compendium of Methods for the Microbiological Examination of Oyedeji, O., Ogunbanwo, S. T., Onilude, A. A. 2013. Predominant Lactic Acid Foods. Fifth edition, American Public Health Association Washington, DC., Bacteria Involved in the Traditional Fermentation of Fufu and Ogi, Two Nigerian USA, p. 769-774. Fermented Food Products. Food and Nutrition Sciences, 04(11), 40–46. Asimi, O. A., Sahu, N. P., Pal, A. K. 2013. Antioxidant activity and antimicrobial https://doi.org/10.4236/fns.2013.411A006 property of some Indian spices. International Journal of Scientific and Research Rojas-Molina, I., Gutiérrez, E., Cortés-Acevedo, M. E., Falcón, A., Bressani, R., Publications, 3(3). ww.ijsrp.org Rojas, A., ... Rodríguez, M. E. (2008). Analysis of Quality Protein Changes in Bello, O. O., Bello, T. K., Amoo, O. T., Atoyebi, Y. O. 2018. Comparative Nixtamalized QPM Flours as a Function of the Steeping Time. Cereal Chemistry evaluation of microbiological and nutritional qualities of various cereal-based Journal, 85(3), 409–416. https://doi.org/10.1094/CCHEM-85-3-0409 paps (Ogi) in Ondo State, Nigeria. International Journal of Environment, Singh, 2018. Recent approaches in food bio-preservation - a review. Open Agriculture and Biotechnology, 3(2), 676–685. Veterinary Journal, 8(1), 104-111. doi: http://dx.doi.org/10.4314/ovj.v8i1.16 https://doi.org/10.22161/ijeab/3.2.49 Stöckheim, A. 2007. J. Lodder (Editor), The Yeasts. A Taxonomic Study. II. Gunaratna, N. S., Moges, D., Groote, H. De. 2019. Biofortified Maize Can überarbeitete Auflage. X und 1385 S., 354 Abb. Amsterdam-London 1970: North Improve Quality Protein Intakes among Young Children in Southern Ethiopia. Holland Publ. Co. $ 65. Zeitschrift Für Allgemeine Mikrobiologie, 12(1), 82–82. Nutrients, 11, 192. https://doi.org/10.3390/nu11010192 https://doi.org/10.1002/jobm.19720120119 Harrigan, W. F. 1998. Laboratory methods in food microbiology. Academic Tandzi, L. N., Mutengwa, C. S., Ngonkeu, E. L. M., Woïn, N., Vernon, G. Press. (2017). Breeding for Quality Protein Maize ( QPM ) Varieties : A Review. Kulamarva, A. G., Sosle, V. R., Raghavan, G. S. V. 2009. Nutritional and Agronomy, 7, 80. https://doi.org/10.3390/agronomy7040080 Rheological Properties of Sorghum. International Journal of Food Properties, Tattari, S., Kota, N., Technical Officer, S., Nimgulkar MPharm Scholar, C.,
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J Microbiol Biotech Food Sci / Olaniran et al. 2020 : 10 (3) 437-443
Polasa, K., Panpatil, V. V, & Nimgulkar, C. (2013). In vitro evaluation on antioxidant and antimicrobial activity of spice extracts of ginger, turmeric and garlic. ~ 143 ~ Journal of Pharmacognosy and Phytochemistry, 2(3), 143–148. http://www.phytojournal.com/archives/2013/vol2issue3/PartC/39.1.pdf Taubeneck, U. 2007. Gibbs B. M, Shapton D. A. (Editors), Identification Methods for Microbiologists. Part B (The Society of Applied Bacteriology, Technical Series No. 2). XI und 212 S., 55 Abb., 37 Tab. London-New York 1968: Academic Press 50 s. Zeitschrift Für Allgemeine Mikrobiologie, 10(3), 232–232. https://doi.org/10.1002/jobm.19700100315 Wakil, S., Ajayi, O. 2013. Production of lactic acid from Starchy-based food substrates. Journal of Applied Biosciences, 71(1), 5673. https://doi.org/10.4314/jab.v71i1.9881 Wood, B. J. B., Holzapfel, W. H. (Editors). 1995. The Genera of Lactic Acid Bacteria. Boston, MA: Springer US. https://doi.org/10.1007/978-1-4615-5817-0
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TECHNOLOGICAL, ANTIOXIDANT, ANTIMICROBIAL AND SENSORY PROFILES OF SELECTED KINDS OF GRAPE OILS
Veronika Juricová1*, Eva Ivanišová2, Július Árvay3, Lucia Godočíková1, Miroslava Kačániová4,5
Address(es): 1Department of Microbiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, SK-949 76, Slovakia. 2Department of Technology and Quality of Plant Products, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, SK-949 76, Slovakia. 3Department of Chemistry, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, SK-949 76, Slovakia. 4Department of Fruit Science, Viticulture and Enology, Faculty of Horticulture and Landscape Engineering, Slovak University of Agriculture, Tr. A. Hlinku 2, SK- 94976 Nitra, Slovakia. 5Department of Bioenergetics, Food Analysis and Microbiology, Institute of Food Technology and Nutrition, University of Rzeszow, Cwiklinskiej 1, 35-601, Rzeszow, Poland.
*Corresponding author: [email protected] doi: 10.15414/jmbfs.2020.10.3.500-504
ARTICLE INFO ABSTRACT
Received 22. 7. 2020 The aim of this study was to determine technological (fat content, acid number, peroxide value, oxidation stability, fatty acid profiles), Revised 10. 9. 2020 antioxidant (DPPH method), antimicrobial activity (antibacterial and antifungal) and sensory profiles of 4 kinds of Slovak grape seed Accepted 22. 10. 2020 oils (commercial, oil from white grape variety, oil from blue grape variety and oil made from grape after alcoholic fermentation). The Published 1. 12. 2020 fat content ranged from 95.86 % (blue grape variety oil) to 98.23 % (commercial oil). The values of acid number ranged from 0.15 mg -1 -1 -1 KOH. g (blue grape variety oil) to 0.90 mg KOH. g (commercial oil) and of peroxide number from 2.75 mmol O2. kg (blue grape variety oil) to 8.91 mmol O . kg-1 (commercial oil). Values of antioxidant activity (DPPH method) ranged from 47.61 mg TEAC. l-1 Regular article 2 (commercial oil) to 75.30 mg TEAC. l-1 (blue grape variety oil) and oxidation stability from 2.75 h (commercial oil) to 8.91 h (blue grape variety oil). The predominant acid in all samples was essential omega-6 fatty acid linoleic acid, which can decrease the cholesterol levels and it is necessary for the activity of cell membranes. Its content differed from 69.43± 0.00 % in commercial oil to 72.75± 0.04 % in blue grape variety oil. Grape oils inhibited microscopic fungi (Candida albicans), gramnegative G- (Yersinia enterocolitica, Salmonella enterica subs. enterica) and grampositive G+ (Bacillus cereus, Staphylococcus aureus subs. aureus) bacteria. Sensory analysis of tested oils was evaluated good, with the best results in commercial oil, especially consistency and overall appearance. The results of this work characterize and accentuate the qualities of edible grape oils, as a nutritionally valuable food for the nutrition of the human organism.
Keywords: oils, fatty acids, antioxidant activity, antimicrobial activity, Rancimat
INTRODUCTION compounds in the human diet because of their effect in reducing the risk of coronary heart diseases and cancer (Konuskan et al., 2018). Fats are one of the main nutrients essential for human nutrition and the proper The objective of this study was to point out and evaluate the technological, DPPH functioning of the body. They fulfill many important functions in the body, they scavenging activity, antimicrobial and sensory properties of various types of are a source of energy, they participate in the construction of cell membranes, grape edible oils. they are a source of fat-soluble vitamins, antioxidants or other biologically active substances (ISEO, 2016). According to the previous findings, fats should account MATERIAL AND METHODS for 30 % to 35 % of the total energy received per day, of which 2/ 3 should be vegetable fats or oils and the remaining 1/ 3 of fats of animal origin (Augustín, Chemicals 2010). Babinská et al. (2012) noted that vegetable oils are characterized by a high content of polyunsaturated fatty acids, of which essential fatty acids play an All the chemicals used were of analytical grade and were purchased from Sigma- important role, which the human body cannot produce alone and therefore must Aldrich (Germany), CentralChem (Slovakia) and Supelco (USA). be taken through diet. Linoleic acid is found in vegetable oils, with a high content of grape oil (Augustín, 2010). The primary source of these essential fatty acids is Material exclusively vegetable oils. The quality of the fats depends mainly on the composition of the fatty acids and their ratio, on the content of biologically active Four samples of edible grape oils were analyzed – commercial oil, oil from white substances, but also on the method of oil production or processing. Virgin or cold grape variety, oil from red grape variety and oil made from grape after alcoholic pressed oils are produced to preserve as many bioactive substances as possible by fermentation. Three samples came from a local producer from the Slovak using a gentle low temperature process (Ogrodowska et al., 2017). However, Republic, which uses grape seeds, forming waste material in wine production, to due to the content of these substances, they are more prone to deterioration, so produce grape oil. All the edible oils analyzed were obtained by "cold pressing" they should be consumed fresh and suitably stored (EFISC Guide, 2014). Grape technology. It is a gentle way of producing edible oils, at a temperature of not seed oils include about poly- and monounsaturated fatty acids as in the amount more than 50 °C, so they are referred to as 'Virgin oils'. of 90% particularly of linoleic acid (58-78%, C18:2) followed by oleic acid (3- 15%, C18:1) and minor amounts of saturated fatty acids (10%). Polyunsaturated fatty acids such as linoleic acid are referred to in the literature as desirable
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Determination of acid number Determination of antibacterial and antifungal properties of edible vegetable oils by selected microorganisms using disc diffusion method The 5 g of sample was weighed into an Erlenmeyer flask, then for each sample, 100 ml of a mixture of equal volumes of ethanol and chloroform were prepared The disc diffusion method is one of the qualitative methods for determining the and neutralized to the indicator phenolphthalein to a slightly pink colour with 3 susceptibility of microbial strains to antimicrobial agents. Depending on the size % ethanolic potassium hydroxide solution. The prepared dissolution system was of the inhibitory zones formed around the disks soaked in the antimicrobial on poured into a sample and the mixture was gently heated to boiling. After mixing the solid soil, the strains can be divided into sensitive or resistant ones. The disc 5 drops of phenolphthalein indicator were added and the contents were titrated diffusion method does not determine the degree of sensitivity of individual with 0.1 mol. l-1 ethanolic potassium hydroxide solution with hot and continuous strains merely to whether the microorganisms are sensitive or resistant to selected stirring until a pinkish colour was observed for 30 s. The assay was performed 3 test substances. A Petri dish filled with Mueller-Hinton agar was used for each times for each sample (Příbela, 1993). bacterial strain. A Petri dish filled with Sabouraud agar was used for yeast strain. Analyzes were done in three replicates. The following strains obtained from the Determination of peroxide number Czech collection of microorganisms (Brno, Czech Republic) were studied: Escherichia coli CCM 3954, Haemophilus influenzae CCM 4454, Klebsiella In a volumetric flask, 75 ml of a 1:1.5 mixture of chloroform and acetic acid was pneumoniae CCM 2318, Yersinia pneumoniae CCM 2318, Salmonella enterica prepared, than 8 g of the sample was weighed into an Erlenmeyer flask and 25 ml subs. enterica CCM 3807, Bacillus cereus CCM 2010, Staphylococcus aureus mixture of chloroform-acetic acid was added. The contents were heated to not subs. aureus CCM 2461, Clostridium perfringens CCM 4435, Streptococcus more than 40 °C to dissolve the sample. Then 1 ml of potassium iodide solution pneumoniae CCM 4501, Candida albicans CCM 8186. was added. The sealed flask was placed in the dark for 20 min. Subsequently, 50 ml of distilled water was added, the contents were mixed; 5 ml of starch oil was Sensory evaluation of oils added and mixed again. The contents of the flask were titrated with sodium thiosulphate solution 0.02 M until discolouration. The test was performed 3 times Sensory analysis of edible grape oils was carried out by twelve evaluators (aged for each sample (Příbela, 1993). 25 to 65; 7 women and 5 men). The panellists (informed laymen) were asked to evaluate the following characteristics: overall appearance, smell (overall), smell Determination of fat content (intensity), foreign smell, taste (overall), taste (intensity), aftertaste, consistency, overall impression after consumption. The assessments were on a 9-point hedonic The fat content of the samples was determined using a fat extractor - scale ranging from 9 (like extremely) to 1 (dislike extremely) for each AncomXT15 (ANKOM Technology, New York, USA) using the manufacturer's characteristic. The results of sensory evaluation from individual evaluators were recommended methodology. Petroleum ether was used as the extraction agent. recorded in a questionnaire and subsequently processed and evaluated in a graph. Each sample was weighed 1.5 g (per cellit) into special bags (XT4 filter bag) and then closed with a melter. Sample bags were dried in an oven (WTB, Binder, Statistical analysis Germany) at 105 °C for 3 hours. Then, they were placed in a desiccator and then weighed. Subsequently, they were placed on an apparatus where extraction with All experiments were carried out in triplicate. The standard deviations for each petroleum ether was carried out at 90 °C for 60 minutes. After extraction, the replication were used for calculation of determined values. The experimental data bags were again dried in an oven (WTB, Binder, Germany) at 105 °C for 30 were subjected to analysis of variance (Duncan's test) at the confidence level of minutes, placed in a desiccator and weighed. The test was performed in two 0.05 (SAS, 2009). replicates. RESULTS AND DISCUSSION Determination of antioxidant activity – DPPH method Determination of acid number The 3,6 ml of DPPH radical (0.025 g was dissolved in 100 ml of ethanol and then diluted as necessary) and 0.4 ml of the sample was pipetted into a cuvette. The Due to the high oil content, the seeds and products of them are subject to mixture was rapidly mixed, centrifuged (VS-100 BN 14000 RPM) and placed in autooxidation. This process is undesirable and causes unpleasant odours and the dark for 10 minutes. Subsequently, the decrease in absorbance on the bitterness of the products. Oxidative and chemical changes in the oil are spectrophotometer (Jenway 6405 UV/ Vis) was monitored at 515 nm. characterized by an increase in acid number (due to an increase in the free fatty Antioxidant activity was expressed on the basis of the Trolox calibration curve acid content of the oil). This parameter indicates the amount of acidic substances (TEAC) in mg TEAC per litre sample (10-100 mg.l-1; R2 = 0.9881). produced by the oxidation of hydrocarbons. This means that the acid number Measurements were repeated 3 times (Sanchez-Moreno et al., 1998). directly determines the degree of oil degradation. Low endpoint values are desirable (Deáková, 2016). The measured values of the acid number of the tested Determination of oxidation stability by Rancimat method oils ranged from 0.15±0.06 to 0.90±0.00 mg KOH.g-1, as can be seen in Table 1. It can be seen from the evaluation of grape oils that all the grape oils tested had The method is based on an accelerated process of oxidation and volatile values below 1 mg KOH.g-1. The highest value of acid number was determined in formation by exposing samples to elevated temperatures with simultaneous air commercial oil and the lowest was determined in blue grape variety oil injection. The time required for measurement is usually several hours instead of (0.15±0.06 mg KOH.g-1). Among the Slovak variety, the highest acid number had weeks or months. The method simulates an accelerated aging process. The a white grape variety oil sample, with a value of 0.64±0.07 mg KOH.g-1. All volatile oxidation products are transferred to the vessel by a stream of air where samples of the tested oils met the requirements for the maximum permissible acid they are absorbed in the measuring solution (distilled water), where the value of the ordinance, up to 4 mg KOH.g-1 for virgin fats and oils. Similar conductivity of the measuring solution is continuously recorded. The results of acid number were also observed in study of Maszewska et al. (2018) – measurement was carried out on a Rancimat 893 (Metrohm, Switzerland), which their obtained values in grapeseed oil ranged from 0.3 to 0.39 mg KOH.g-1. works and records the entire measurement process using the Stabnet 893 software. The measurement results are a software curves with induction time. Table 1 The results of acid number, peroxide number and fat content of analyzed samples Determination of fatty acid methyl esters content by GC-FID method Peroxide Acid number Fat content Sample -1 number [mg KOH. g ] -1 [%] Prepared, esterified oil samples should be diluted 1: 19 (50 µl FAME + 950 µl N- [mmol O2. kg ] hexane) before analysis. The qualitative and quantitative determination of FAME Commercial oil 0.90±0.11a 8.91±1.05a 98.23±2.32a was performed by gas chromatography detection on a flame ionization detector White grape 0.64±0.02a 6.00±0.93b 96.00±2.49a on an AGILENT 7890B (Agilent Technologies USA). The 1 µl of the diluted variety oil sample was injected with the CombiPAL autosampler into the instrument. Blue grape 0.15±0.03a 2.75±0.14c 95.86±1.68a Separation of fatty acid methyl esters was performed on a HP-88 GC capillary variety oil column (Agilent Technologies USA, 60 m x 0.25 mm x 0,20 µm). All analytical Grape oil after 0.56±0.12a 3.75±0.11c 97.46±1.33a gases used (He, N2, H2, synthetic air) had a purity of 5.0. Detection of separated fermentation components was performed using a flame ionization detector. The data was KOH – potassium hydroxide; mean ±standard deviation; different letters in column denote processed online using Agilent OpenLab ChemStation.Standardization and mean values that statistically differ one from another instrument calibration was performed using a 37-component Supelco 37 Component FAME Mix (CRM, TraceCERT, Supelco USA). The qualitative Determination of peroxide number determination of chromatographic analyzes by GC-Fit method was performed by Agilent 5977A MSD mass spectrometry. The data obtained were compared with The peroxide number is another indicator of lipid oxidation or yellow oil the spectral data of the NIST 14.L. evaluation. It characterizes the amount of peroxides and hydroperoxides formed by the action of air oxygen. Low value predetermines good stability of the sample
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against oxidative damage (Deáková, 2016). Parry et al. (2004) reported that the Table 2 The results of antioxidant activity and oxidation stability of analyzed peroxide number and other indicators of the oxidation state of oils are highly samples dependent on the oil production method, which has been demonstrated in a study Antiradical Oxidation stability using a dry method of extracting grape seed oil. Oils obtained by microwave or Sample activity -1 -1 [h] dry heating had a much higher peroxide value (5.6 mmol O2.kg ) than those [mg TEAC. l ] -1 obtained by cold pressing (1.9 mmol O2.kg ). When evaluating grape oils (Table Commercial oil 47.61±0.21d 8.91±0.32a 1), it can be seen significant differences in peroxide value for each samples -1 White grape variety oil 69.05±0.09b 6.00±0.04b ranging from 2.75±0.50 to 8.91±1.01 mmol O2.kg . Commercial oil has a -1 significantly higher value than Slovak grape oils, 8.91±1.01 mmol O2.kg . Blue grape variety oil 75.30±0.15a 2.75±0.09d Among the Slovak samples, the lowest value has the blue grape variety oil Grape oil after fermentation 65.03±1.08c 3.75±0.14c -1 (2.75±0.50 mmol O2.kg ), the highest value, up to twice as high as blue grape TEAC – Trolox equivalent antioxidant capacity; mean± standard deviation; different letters -1 variety oil, has the white grape variety oil sample (6.00±0.25 mmol O2.kg ). All in column denote mean values that statistically differ one from another samples examined meet the requirements of the decree, which stipulates a -1 peroxide value of not more than 15 mmol O2.kg for virgin fats and oils. In study Determination of oxidation stability by Rancimat method of Maszewska et al. (2018) peroxide number in grapeseed oil ranged from 2.6 to -1 3 mmol O2.kg , which is comparable with our results in case of blue grape Dyerberg et al. (2010) reported that unsaturated fats in fats are readily oxidized. variety and grape oil after fermentation. Oxidation products such as hydroperoxides and their breakdown products (aldehydes, ketones, organic acids) cause unwanted chemical or sensory changes Determination of the amount of fat in fat products, and the consumption of oxidized fats poses a health risk (free radical formation, lipoprotein oxidation, mutagenic changes in the body). As we can see in Table 1, the fat content of the grape oil samples ranged from Protection against oxidation consists in eliminating the adverse effects of fat 95.86±1.68 to 98.23±2.32 %. Significant differences are visible in grape oils – oxidation. When comparing grape oils (Table 2), we can conclude that Slovak commercial oil has the highest content of 98.23±2.32 %, while the lowest has samples achieved lower values of oxidation stability than commercial oil. Slovak blue grape variety oil of 95.86±1.68 %. Among Slovak oils, the highest fat samples reached values from 2.75±0.09 h to 6.00±0.04 h, while commercial oil content has grape oil after fermentation. Since the samples analyzed are cold reached the highest value of stability – 8.91±0.32 h. Among the Slovakian, the pressed oils without refining, it is evident that these oils contain dyes, fiber, longest induction time has reached white grape variety oil. proteins - therefore the fat content is not 100 %. Generally the fastest oxidation of grapeseed oil can be explained by fatty acid Moreau et al. (2009) reported that pigments, such as carotenoids and composition. Grape oil contained approximately 68% polyunsaturated acids; chlorophylls, present in unrefined oils can reduce oil stability, although they can while for example peanut and rapeseed oils contained the smallest amounts of also be a positive attribute to provide an attractive colour. polyunsaturated acids (about 25–28%, respectively) (Maszewska et al., 2018). Oxidative stability of oils is also influenced by antioxidants. The content of Determination of antiradical activity – DPPH method sterols in vegetable oils is from 70 to 1100 mg.100 g-1 of oil. Most of these compounds are found in corn oil, then slightly less in rapeseed oil γ-oryzanol in When comparing the antiradical activity of each type of grape oil, we observed rice bran and tocotrienols and tocopherols in grapeseed and rice bran oils (Ratisz considerable differences. Slovakian species achieved significantly higher values et al. 2016). compared to the commercial oil, which reached 47.61±0.01 mg TEAC.l-1. The antiradical activity values of all samples are shown in Table 2. According to Du Determination of fatty acid methyl esters content by GC-FID method Toita et al. (2001) the DPPH method is considered one of the basic methods for assessing the antiradical activity of pure substances or solutions. The essence is The predominant acid in all samples was linoleic acid (Table 3). Its content the reaction of the test substance with a stable diphenylpicrylhydrazyl radical differed from 69.43± 0.00 % in commercial oil to 72.75± 0.04 % in blue grape (DPPH), in which the radical is reduced in reaction to give DPPH-H variety oil. Elaidic acid was the least represented in the case of white grape (diphenylpicrylhydrazine). This reaction is most often observed variety oil and grape oil after fermentation. In commercial oil was identified spectrophotometrically, measuring absorbance at 515 nm. Konuskan et al. linolelaidic acid in an amount of 0.64± 0.00 %, which was also the least (2018) tested antioxidant activity of cold pressed grape oil (from variety Sirah, represented acid in this oil sample and which was not found in the other grape oil Merlot, Sangiovese, Cabernet Sauvignon and Sauvignon Blanc) by DPPH samples. Equally, essential α-linolenic acid was identified in the blue grape method. Higher activity was determined in their study in oils from blue grape variety oil at 0.43±0.00 %, while in other samples it was not identified. Other varieties with the best results in oil from variety Merlot (17.94%) following by acids found in grape oils without major differences were oleic acid, palmitic acid variety Cabernet Sauvignon (15.32), Sirah (13.28%) and Sangiovese (10.41%). and stearic acid. When comparing the oils from the point of view of origin, we The lowest value was determined in oil from white grape variety Sauvignon can say that there are no significant differences in the content of individual types Blanc (7.04%). of fatty acids. However, linolelaidic acid, which was not found in Slovak samples, is also present in commercial oil. In study of Maszewska et al., (2018) corn and grapeseed oils contained 28.8% and 18.8% of monoenic acids. In their study was also published that the largest share of polyene acids were found in grapeseed oil (68.4%). From a nutritional point of view, the content of polyene fatty acids in oils is a desirable feature. On the other hand, oils of this composition are not very resistant to external factors, they are very susceptible to oxidation e.g. α-linolenic acid. There is concern that long-term consumption of large amounts of this acid can increase cancer risk (Hart et al., 2009).
Table 3 The results of fatty acid methyl esters content of analyzed samples White variety grape oil Blue variety grape oil Grape oil after fermentation Commercial oil Sample [%] [%] [%] [%] C16: 0 8.43±0.02a 7.81±0.05b 7.66±0.02b 7.48±0.03b C18: 0 3.83±0.00a 3.96±0.00a 3.73±0.01b 3.87±0.01a C18: 1n9c 17.35±0.00a 14.34±0.01b 15.55±0.00b 17.84±0.02a C18: 1n9t 0.83±0.00a 0.70±0.00b 0.78±0.00b 0.75±0.00b C18: 2n6t 0.00±0.00b 0.00±0.00 0.00±0.00b 0.64±0.00a C18: 2n6c 69.57±0.02b 72.75±0.04a 72.28±0.01a 69.43±0.00b C18: 3n3c 0.00±0.00b 0.43±0.00a 0.00±0.00b 0.00±0.00b mean ± standard deviation; C16:0=palmitic acid, C18:0=stearic acid, C18:1n9c=oleic acid, C18:1n9t=elaidic acid, C18:2n6t=linolelaidic acid, C18:2n6c=linoleic acid, C18:3n3c=α-linolenic acid; different letters in column denote mean values that statistically differ one from another
Determination of antibacterial and antifungal properties of edible vegetable facultatively anaerobic, cocoid bacteria, the Micrococcaceae family. It occurs on oils by selected microorganisms using disc diffusion method the skin and mucous membranes of humans and animals. Staphylococcus aureus may be responsible for food poisoning due to the consumption of contaminated Bacillus cereus is an aerobic or facultatively anaerobic, G+, rod-shaped bacterium foods such as meat and meat products, eggs, salads, bakery and dairy products sporulating from the Bacillaceae family that occurs in soil and water (Cammack (IFIS, 2009). Grape seed oil has a toxicity effect on some pathogens, suggesting et al., 2006). Staphylococcus aureus subs. aureus belongs to the genus G+, an antimicrobial feature. In fact, the oil extracted from grape seeds had an
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inhibitory effect on the growth of Staphylococcus aureus and Escherichia coli findings suggest that the use of resveratrol would aid traditional therapies in (Baydar et al., 2006; Rotava et al., 2009). The antimicrobial activity displayed which antibiotics are ineffective (Subramanian et al., 2014). Yersinia by phenolic compounds, such as resveratrol, involves the induction of oxidative enterocolitica belongs to the family of facultatively anaerobic, rod-shaped, G- damage to bacterial membrane, especially E. coli, without affecting the host cells. bacteria of the Enterobacteriaceae family. It occurs in soil and water, in the These findings suggest that the use of resveratrol would aid traditional therapies gastrointestinal tract of animals (such as pigs and rodents). Some species are able in which antibiotics are ineffective (Subramanian et al., 2014). Bacillus cereus to survive and reproduce even at low temperatures, so they can pose a danger to is an aerobic or facultatively anaerobic, G+, rod-shaped bacterium sporulating refrigerant foods (IFIS, 2009). Salmonella enterica subs. enterica belongs to from the Bacillaceae family that occurs in soil and water (Cammack et al., facultative anaerobic, rod-shaped, G-bacteria, Enterobacteriaceae. It occurs 2006). Staphylococcus aureus subs. aureus belongs to the genus G+, facultatively mainly in soil, water and food (eg raw meat and seafood, eggs, dairy products) anaerobic, cocoid bacteria, the Micrococcaceae family. It occurs on the skin and and in the gastrointestinal tract of humans and animals (especially poultry and mucous membranes of humans and animals. Staphylococcus aureus may be pigs) (Cammack et al., 2006). responsible for food poisoning due to the consumption of contaminated foods Candida albicans belongs to the yeast genus, the class of Saccharomyces. It such as meat and meat products, eggs, salads, bakery and dairy products (IFIS, occurs in soil and on plants (IFIS, 2009). Palma et al. (1999) carried out a study, 2009). Grape seed oil has a toxicity effect on some pathogens, suggesting an where grape seeds were exerted the strongest antimicrobial action against S. antimicrobial feature. In fact, the oil extracted from grape seeds had an inhibitory coagulans E.cloacae, C. freundii and E.coli. The later bacteria were sensitive to effect on the growth of Staphylococcus aureus and Escherichia coli (Baydar et all tested concentrations of fraction A, even the lowest one. S. aureus was only al., 2006; Rotava et al., 2009). The antimicrobial activity displayed by phenolic moderately sensitive to the highest concentration. B. cereus and the fungus A. compounds, such as resveratrol, involves the induction of oxidative damage to flavus were resistant to fraction A at every concentration bacterial membrane, especially E. coli, without affecting the host cells. These
Table 4 The results of antimicrobial activity against G+ [mm] Staphylococcus aureus Clostridiun Streptococcus Bacillus cereus Sample subs. aureus perfringens pneumoniae CCM 2010 CCM 2461 CCM 4435 CCM 4501 Commercial oil 4.33±2.08ab 5.33±0.53a 3.00±1.73a 0.10±0.01b White grape variety 3.67±2.08ab 2.33±0.58b 2.67±0.58a 0.00±0.00c oil Blue grape variety oil 6.67±0.58a 4.67±0.58a 2.00±0.00a 0.00±0.00c Grape oil after 2.67±0.58b 4.33±0.58a 2.33±0.58a 0.13±0.03a fermentation mean ±standard deviation; different letters in column denote mean values that statistically differ one from another
Table 5 Results of antimicrobial activity against G- [mm] Klebsiella Haemophilus Salmonella enterica Yersinia enterocolitica Escherichia coli Sample pneumoniae influenzae subs. enterica CCM 5671 CCM 3954 CCM 2318 CCM 4456 CCM 3807 Commercial oil 2.33±0.58a 2.67 ±1.15bc 0.03±0.06c 2.67±1.15a 0.13±0.01a White grape 1.67±0.58a 2.00±0.00c 0.00±0.00c 4.00±1.73a 0.03±0.01c variety oil Blue grape 2.33±0.58a 3.00±1.00ab 0.07±0.01b 2.00±1.00a 0.00±0.00d variety oil Grape oil after 1.67±0.58a 4.33±0.58a 0.17±0.02a 2.67±0.58a 0.07±0.12b fermentation mean ±standard deviation; different letters in column denote mean values that statistically differ one from another
Table 6 Results of antifungal activity [mm] White variety grape oil Blue variety grape oil Grape oil after fermentation Commercial oil Sample
Candida albicans 0.10±0.01a 0.0±0.00b 0.03±0.01b 0.10±0.0.01a CCM 8186 mean ±standard deviation; different letters in column denote mean values that statistically differ one from another
Sensory evaluation of oil
We can conclude that the overall appearance was evaluated for all samples with high values. The worst rating was grape oil after fermentation with a value of 7.15. The presence of a foreign smell was most present in grape oil after fermentation a 6.00 point. From the point of view of consistency, all oils achieved a high rating from 7.23 to 8.31 points, the highest value attributed to white grape variety oil. Blue grape variety oil was characterized by its dark green colour and typical grape aroma and taste. There was also a foreign smell characterized as being gently beer, acidic, to rubber after oxidation. The oil also left a sour, beer or metal aftertaste. White grape variety oil was light green colour, alcoholic, wine odour. scent of mussels and herbs. In addition to the grapes, the taste was slightly soft and the wine was slightly scraped in the throat. Grape oil after fermentation, had a dark Figure 1 Sensory evaluation of oil green colour, characterized by a wine, acetic, ethanol or alcohol flavour, similar to that of the B-complex. The aftertaste has been characterized as alcoholic, Commercial oil smelled of the presence of foreign smells of alcohol or fermented liqueur, vinous, drug-like or nail polish, characterized by consumers as being grapes. The scent was, according to the evaluators, unpleasant, acidic, burgundy unsuitable for oil. or yeast, and some had a balsamic vinegar smell. The evaluators noted slightly alcoholic, yeast to sweetish aftertaste that caused sore throat. The colour was gently green. Colour is a qualitative property that significantly affects the consumer's oil rating. Green pigments, in particular chlorophyll content, impart undesirable colour to
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berry plant vegetable oils and can also promote oxidation thereof (Gutiérrez et. Pharmaceutical Education and Research, 53(1), 144–150. al., 2007). https://doi.org/10.5530/ijper.53.1.19. Moreau, R. A., & Kamal-Eldin, A. (2009). Gourmet and Health-Promoting CONCLUSION Specialty Oils, AOCS Press. Maszewska, M., Florowska, A., Dłużewska, E., Wroniak, M., Marciniak- The results of our study point to a higher quality of local Slovak types of grape Lukasiak, K., & Żbikowska, A. (2018). Oxidative Stability of Selected Edible oils compared to commercial oil, the reason could be higher quality of raw Oils. Molecules, 23(7), 1746. https://doi.org/10.3390/molecules23071746. material, well-chosen variety of crop or harvesting method. The quality of grape Ogrodowska, D., Tańska, M., & Brandt, W. (2017). The Influence of Drying oils is characterized by very low acidity values, as an important indicator of fat Process Conditions on the Physical Properties, Bioactive Compounds and and oil status, the lowest measured value of 0.15 ± 0.06 mg KOH. g-1 in blue Stability of Encapsulated Pumpkin Seed Oil. Food and Bioprocess Technology, grape variety oil. The peroxide number characterizing the content of peroxides 10(7), 1265–1280. https://doi.org/10.1007/s11947-017-1898-z. and hydroperoxides formed by the undesirable oxidation process, which Palma, M., Taylor, L. T., Varela, R. M., Cutler, S. J., & Cutler, H. G. (1999). negatively affects the quality or nutritional value of the oil, was characterized by Fractional Extraction of Compounds from Grape Seeds by Supercritical Fluid significant differences between the grape variety of oil, values from 2.75 ± 0.50 Extraction and Analysis for Antimicrobial and Agrochemical Activities. Journal -1 -1 mmol O2. kg (blue grape variety oil) to 6.00 ± 0.25 mmol O2. kg (white grape of Agricultural and Food Chemistry, 47(12), 5044–5048. variety oil). The antioxidant activity of the oils was measured by the DPPH https://doi.org/10.1021/jf990019p. radical with which the antioxidants present in the oil reacted. The highest value Parry, J., & Yu, L. (2006). Fatty Acid Content and Antioxidant Properties of of 75.30± 0.00 mg TEAC. l-1 was achieved in blue grape variety oil. With the Cold-pressed Black Raspberry Seed Oil and Meal. Journal of Food Science, GC-FID method, have been determined the fatty acid content of individual oil 69(3), FCT189–FCT193. https://doi.org/10.1111/j.1365-2621.2004.tb13356.x. samples and the measurement is also interesting from the point of view of the Příbela, A. (1993). Analýza potravín. Bratislava: STU, 2., 394. presence of essential fatty acids. Omega-6 fatty acid linoleic had the highest Ratusz, K., Popis, E., Ciemniewska-Żytkiewicz, H., & Wroniak, M. (2016). proportion in blue grape variety oil at 72.75 ± 0.04 % and α-linolenic acid was Oxidative stability of camelina (Camelina sativa L.) oil using pressure also recorded. In commercial oil omega-6 fatty acid linoelaide was present, differential scanning calorimetry and Rancimat method. Journal of Thermal which was not present in another oil samples. The measurements also confirmed Analysis and Calorimetry, 126(1), 343–351. https://doi.org/10.1007/s10973-016- the anibacterial and antifungal properties of vegetable oils. The greatest 5642-0. inhibition in G+ bacteria was recorded on Bacillus cereus, in blue grape variety Rotava, R., Zanella, I., & Da Silva, L.P. (2009). Antibacterial, antioxidant and oil. In sensory evaluation, the best overall impression was achieved by the white tanning activity of grape by product. Ciencia Rural, 39, 941–944. grape variety oil and the least points by the blue grape variety oil. All tested SAS. Users Guide Version 9.2. SAS/STAT (r). SAS Institute Inc. Cary, NC, grape oil samples comply with the requirements of the Food Code of the Slovak USA. 2009. Republic. Sánchés-Moreno, C., Larrauri, A., & Saura-Calixto, F.A. (1998). The procedure to measure the antioxidant efficiency of polyphenols. Journal of the Science of Acknowledgments: Work was supported by Research Center AgroBioTech built Food and Agriculture, 76, 270–276. https://doi.org/10.1002/(sici)1097- in accordance with the project Building Research Center „AgroBioTech“ ITMS 0010(199802)76:2<270::aid-jsfa945>3.0.co;2-9. 26220220180. Subramanian, M., Goswami, M., Chakraborty, S., & Jawali, N. (2014). Resveratrol induced inhibition of Escherichia coli proceeds via membrane REFERENCES oxidation and independent of diffusible reactive oxygen species generation. Redox Biology, 2, 865–872. https://doi.org/10.1016/j.redox.2014.06.007. Augustín, J. (2010). Plant and Animal Oils and Technical Oils. Epicure, 4(17), 40-49. Babinská, K., & Béderová, A. (2012). The Importance of Fats and Their Consumption in Population SR. Bratislava: Institute of Physiology LF UK, 32- 38. Baydar, N. G., Sagdic, O., Ozkan, G., & Cetin, S. (2006). Determination of antibacterial effects and total phenolic contents of grape (Vitis viniferaL.) seed extracts. International Journal of Food Science & Technology, 41(7), 799–804. https://doi.org/10.1111/j.1365-2621.2005.01095.x. Cammack, R., Attwood, T. K., Campbell, P. N., Parish, J. H., Smith, A. D., Stirling, J. L., & Vella, F. (2006). Oxford Dictionary of Biochemistry and Molecular Biology. New York: Oxford University Press, 2. Deáková, Ľ. (2016). Chemické zloženie a oxidačná stabilita semien a oleja maku siateho (Papaver somniferum L.). Mak siaty pre Slovensko Piešťany : NPPC, VÚRV, 28-29. Du Toit, R., Volsteedt, Y., & Apostolides, Z. (2001). Comparison of the antioxidant content of fruits, vegetables and teas measured as vitamin C equivalents. Toxicology, 166(1-2), 63–69. https://doi.org/10.1016/s0300- 483x(01)00446-2. Dyerberg, J., Madsen, P., Møller, J. M., Aardestrup, I., & Schmidt, E. B. (2010). Bioavailability of marine n-3 fatty acid formulations. Prostaglandins, Leukotrienes and Essential Fatty Acids, 83(3), 137–141. https://doi.org/10.1016/j.plefa.2010.06.007. EFISC GUIDE (2014). Sector reference document on the manufacturing of safe feed materials from oilseed crushing and vegetable oil refining. Brussels: EFISC Aisbl., 162. Gutiérrez, F., Jímenez, B., Ruíz, A., & Albi, M. A. (1999). Effect of Olive Ripeness on the Oxidative Stability of Virgin Olive Oil Extracted from the Varieties Picual and Hojiblanca and on the Different Components Involved. Journal of Agricultural and Food Chemistry, 47(1), 121–127. https://doi.org/10.1021/jf980684i. IFIS (2009!. Dictionary of Food Science and Technology. Singapur : IFIS Publishing - International Food Information Service, 2. Hart, A., Tjonneland, A., Olsen, A., Overvad, K., Bergmann, M.M., Boeing, H., Nagel, G., Linseisen, J., HallmansS, G., & Danielsson, Å. (2009). Linoleic acid, a dietary n-6 polyunsaturated fatty acid, and the aetiology of ulcerative colitis: a nested case-control study within a European prospective cohort study. Gut, 58(12), 1606–1611. https://doi.org/10.1136/gut.2008.169078. INSTITUTE OF SHORTENING AND EDIBLE OILS, INC, ISEO. (2016). Food fats and oils. Washington, DC. 3. Konuskan, D. B., Kamiloglu, O., & Demirkeser, O. (2019). Fatty Acid Composition, Total Phenolic Content and Antioxidant Activity of Grape Seed Oils Obtained by Cold- Pressed and Solvent Extraction. Indian Journal of
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